Forskolin Cancer 09 22 2017

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1. Improved differentiation method

By Clevers, Johannes Carolus; Gehart, Helmuth
From PCT Int. Appl. (2017), WO 2017149025 A1 20170908, Language: English, Database: CAPLUS
The invention relates to differentiation methods for progenitor cells, e.g. mammalian epithelial stem cells, differentiation
media for use in said methods, organoids and cells obtainable by said methods and uses, including therapeutic uses,
thereof.
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Copyright © 2017 American Chemical Society (ACS). All Rights Reserved.
2. Synthesis of novel forskolin isoxazole derivatives with potent anti-cancer activity against breast cancer cell
lines
By Burra, Srinivas; Voora, Vani; Rao, Ch. Prasad; Vijay Kumar, P.; Kancha, Rama Krishna; David Krupadanam, G. L.
From Bioorganic & Medicinal Chemistry Letters (2017), 27(18), 4314-4318. Language: English, Database: CAPLUS,
DOI:10.1016/j.bmcl.2017.08.033
Forskolin C1-isoxazole derivs. (3,5-regioisomers) were synthesized regioselectively by adopting 1,3-dipolar cycloaddns.
These derivs. were tested using estrogen receptor pos. breast cancer cell lines MCF-7 and BT-474. A majority of the
compds. exhibited activity against the p53-pos. MCF-7 breast cancer cells but not against the p53-neg. BT-474 breast
cancer cells. Among forskolin derivs., compds. I [R3 = 4-MeOC6H4, 3,4,5-(MeO)3C6H2], II [R1 = H, R2 = COMe, R3 = 4-
MeOC6H4, 4-PhCH2OC6H4, 2,4,6-(MeO)3C6H2, Ph; R1 = COMe, R2 = H, R3 = 2,4,6-(MeO)3C6H2], III [R1 = H, R2 =
COMe, R3 = 3,4-(MeO)2C6H3; R1 = COMe, R2 = H, R3 = 3,4-(MeO)2C6H3] exhibited higher anti-cancer activity against
MCF-7 cell line with an IC50 ≤ 1 µM. The deriv. II (R1 = H, R2 = COMe, R3 = 4-PhCH2OC6H4) exhibited the highest
activity in both p53-pos. (MCF-7) and p53-neg. (BT-474) breast cancer cell lines with an IC50 of 0.5 µM.
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3. Antihelminthic drug niclosamide inhibits CIP2A and reactivates tumor suppressor protein phosphatase 2A in
non-small cell lung cancer cells
By Kim, Myeong-ok; Choe, Min Ho; Yoon, Yi Na; Ahn, Jiyeon; Yoo, Minjin; Jung, Kwan-Young; An, Sungkwan; Hwang,
Sang-Gu; Oh, Jeong Su; Kim, Jae-Sung
From Biochemical Pharmacology (Amsterdam, Netherlands) (2017), Ahead of Print. Language: English, Database:
CAPLUS, DOI:10.1016/j.bcp.2017.08.009
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Protein phosphatase 2A (PP2A) is a crit. tumor suppressor complex responsible for the inactivation of various
oncogenes. Recently, PP2A reactivation has emerged as an anticancer strategy. Cancerous inhibitor of protein
phosphatase 2A (CIP2A), an endogenous inhibitor of PP2A, is upregulated in many cancer cells, including non-small cell
lung cancer (NSCLC) cells. We demonstrated that the antihelminthic drug niclosamide inhibited the expression of CIP2A
and reactivated the tumor suppressor PP2A in NSCLC cells. We performed a drug-repurposing screen and identified
niclosamide as a CIP2A suppressor in NSCLC cells. Niclosamide inhibited cell proliferation, colony formation, and tumor
sphere formation, and induced mitochondrial dysfunction through increased mitochondrial ROS prodn. in NSCLC cells;
however, these effects were rescued by CIP2A overexpression, which indicated that the antitumor activity of niclosamide
was dependent on CIP2A. We found that niclosamide increased PP2A activity through CIP2A inhibition, which reduced
the phosphorylation of several oncogenic proteins. Moreover, we found that a niclosamide analog inhibited CIP2A
expression and increased PP2A activity in several types of NSCLC cells. Finally, we showed that other well-known
PP2A activators, including forskolin and FTY720, did not inhibit CIP2A and that their activities were not dependent on
CIP2A. Collectively, our data suggested that niclosamide effectively suppressed CIP2A expression and subsequently
activated PP2A in NSCLC cells. This provided strong evidence for the potential use of niclosamide as a PP2A-activating
drug in the clin. treatment of NSCLC.
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4. Differential proteome expression analysis of androgen-dependent and -independent pathways in LNCaP

prostate cancer cells
By Cha, Seho; Shin, Dong Hoon; Seok, Jun Ryeong; Myung, Jae Kyung
From Experimental Cell Research (2017), 359(1), 215-225. Language: English, Database: CAPLUS,
DOI:10.1016/j.yexcr.2017.07.026
Prostate cancer (PC) is one of the leading causes of cancer death in men. It commonly develops in older males, but the
no. of younger men diagnosed with the disease has increased in recent years. Hormone therapies, such as chem. and
surgical methods that inhibit androgen synthesis or androgen receptor (AR) activation, have been used for advanced
disease. However, castration-resistant PC (CRPC), which exhibits androgen-independent mechanisms for activating AR,
develops after a few years of such treatment and no therapy is available. In this study, we examd. differences in the
proteomes assocd. with the androgen-dependent (DHT treatment) and -independent (FSK, forskolin treatment) AR
signaling conditions in LNCaP prostate cancer cells. Moreover, we used EPI-001, which inhibits AR-mediated
transcriptional activity, to examine whether the obsd. differences in protein expression were directly affected by AR-
mediated mechanisms. A total of 213 protein spots were matched in our 2-dimensional gel electrophoresis (2DE) anal.
and 8 proteins with significant expression changes in our 5 different treatment groups were identified by mass
spectrometry. Among these proteins, expression levels of PEPCK-M, catalase, tubulin alpha chain, alpha-enolase, and
endoplasmic reticulum resident protein 29 were significantly altered by DHT and the levels of HSP 90 and EF-Tu were
changed by FSK. These changes were blocked by EPI-001 in DHT-regulated proteins, PEPCK-M, catalase, and tubulin
alpha chain and FSK-regulated EF-Tu protein. The results from our immunohistochem. anal. using in vivo xenograft
models were consistent with the 2DE data. We therefore report the first identification of differences in proteins that are
significantly regulated under androgen-dependent and -independent AR signaling conditions. Our findings could suggest
a possible mol. mechanism through which AR is activated in the absence and/or presence of androgen, and might help
explain the inhibitory action of EPI-001 on CRPC.
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5. Global alteration of the drug-binding pocket of human P-glycoprotein (ABCB1) by substitution of fifteen
conserved residues reveals a negative correlation between substrate size and transport efficiency
By Vahedi, Shahrooz; Chufan, Eduardo E.; Ambudkar, Suresh V.
From Biochemical Pharmacology (Amsterdam, Netherlands) (2017), Ahead of Print. Language: English, Database:
CAPLUS, DOI:10.1016/j.bcp.2017.07.014
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P-glycoprotein (P-gp), an ATP-dependent efflux pump, is linked to the development of multidrug resistance in cancer
cells. However, the drug-binding sites and translocation pathways of this transporter are not yet well-characterized. We
recently demonstrated the important role of tyrosine residues in regulating P-gp ATP hydrolysis via hydrogen bond
formations with high affinity modulators. Since tyrosine is both a hydrogen bond donor and acceptor, and non-covalent
interactions are key in drug transport, in this study we investigated the global effect of enrichment of tyrosine residues in
the drug-binding pocket on the drug binding and transport function of P-gp. By employing computational anal., 15
conserved residues in the drug-binding pocket of human P-gp that interact with substrates were identified and then
substituted with tyrosine, including 11 phenylalanine (F72, F303, F314, F336, F732, F759, F770, F938, F942, F983,
F994), two leucine (L339, L975), one isoleucine (I306), and one methionine (M949). Characterization of the tyrosine-rich
P-gp mutant in HeLa cells demonstrated that this major alteration in the drug-binding pocket by introducing fifteen addnl.
tyrosine residues is well tolerated and has no measurable effect on total or cell surface expression of this mutant.
Although the tyrosine-enriched mutant P-gp could transport small to moderate size (<1000 Daltons) fluorescent
substrates, its ability to transport large (>1000 Daltons) substrates such as NBD-cyclosporine A, Bodipy-paclitaxel and
Bodipy-vinblastine was significantly decreased. This was further supported by the physico-chem. characterization of
seventeen tested substrates, which revealed a neg. correlation between drug transport and mol. size for the tyrosine-
enriched P-gp mutant.
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6. Assay of Calcium Transients and Synapses in Rat Hippocampal Neurons by Kinetic Image Cytometry and
High-Content Analysis: An In Vitro Model System for Postchemotherapy Cognitive Impairment
By McDonough, Patrick M.; Prigozhina, Natalie L.; Basa, Ranor C. B.; Price, Jeffrey H.
From Assay and Drug Development Technologies (2017), 15(5), 220-236. Language: English, Database: CAPLUS,
DOI:10.1089/adt.2017.797
Postchemotherapy cognitive impairment (PCCI) is commonly exhibited by cancer patients treated with a variety of
chemotherapeutic agents, including the endocrine disruptor tamoxifen (TAM). The etiol. of PCCI is poorly understood.
Our goal was to develop high-throughput assay methods to test the effects of chems. on neuronal function applicable to
PCCI. Rat hippocampal neurons (RHNs) were plated in 96- or 384-well dishes and exposed to test compds. (forskolin
[FSK], 17β-estradiol [ES]), TAM or fulvestrant [FUL], aka ICI 182,780) for 6-14 days. Kinetic Image Cytometry® (KIC®)
methods were developed to quantify spontaneously occurring intracellular calcium transients representing the activity of
the neurons, and high-content anal. (HCA) methods were developed to quantify the expression, colocalization, and
puncta formed by synaptic proteins (postsynaptic d. protein-95 [PSD-95] and presynaptic protein Synapsin-1 [Syn-1]).
As quantified by KIC, FSK increased the occurrence and synchronization of the calcium transients indicating stimulatory
effects on RHN activity, whereas TAM had inhibitory effects. As quantified by HCA, FSK also increased PSD-95 puncta
and PSD-95:Syn-1 colocalization, whereas ES increased the puncta of both PSD-95 and Syn-1 with little effect on
colocalization. The estrogen receptor antagonist FUL also increased PSD-95 puncta. In contrast, TAM reduced Syn-1
and PSD-95:Syn-1 colocalization, consistent with its inhibitory effects on the calcium transients. Thus TAM reduced
activity and synapse formation by the RHNs, which may relate to the ability of this agent to cause PCCI. The results
illustrate that KIC and HCA can be used to quantify neurotoxic and neuroprotective effects of chems. in RHNs to
investigate mechanisms and potential therapeutics for PCCI.
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7. Use of low molecular weight compounds for treatment of glioblastoma

By Ha, Yun; Oh, Jin Su; Kim, Yong Bo; Cha, Ryeo Hwa
From Repub. Korean Kongkae Taeho Kongbo (2017), KR 2017080390 A 20170710, Language: Korean, Database:
CAPLUS
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The present invention relates to the therapeutic use of low mol.
wt. compds. for glioblastoma. More particularly, the present
invention relates to a method for selectively inhibiting the
proliferation of neuroblastoma and converting neuroblastoma
into neuronal cells using only low mol. compds. CHIR99021
and Forskolin. The present invention suggests a next-
generation treatment of a new paradigm that directly converts
the fate of malignant glioblastoma to benign neurons, unlike the
existing treatment principle for neoplastic tumors. The compd.
according to the present invention is a customized therapeutic
compn. for a glioblastoma, and unlike conventional anticancer
drug therapy and surgical operation, it can be used for
selecting a neuroblastoma cell line without side effects such as
development of resistant mutant strains, recurrence by tumor
stem cells Which can effectively treat a glioblastoma cell line by
inhibiting proliferation or converting to neuronal cells.
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8. Multiple SLC and ABC transporters contribute to the placental transfer of entecavir
By Ma, Zhiyuan; Yang, Xi; Jiang, Ting; Bai, Mengru; Zheng, Caihong; Zeng, Su; Sun, Dongli; Jiang, Huidi
From Drug Metabolism & Disposition (2017), 45(3), 269-278. Language: English, Database: CAPLUS,
DOI:10.1124/dmd.116.073304
Entecavir (ETV), a nucleoside analog with high efficacy against hepatitis B virus, is recommended as a first-line antiviral
drug for the treatment of chronic hepatitis B. However, scant information is available on the use of ETV in pregnancy.
To better understand the safety of ETV in pregnant women, we aimed to demonstrate whether ETV could permeate
placental barrier and the underlying mechanism. Our study showed that small amt. of ETV could permeate across
placenta in mice. ETV accumulation in activated or nonactivated BeWo cells (treated with or without forskolin) was
sharply reduced in the presence of 100 µM of adenosine, cytidine, and in Na+ free medium, indicating that nucleoside
transporters possibly mediate the uptake of ETV. Furthermore, ETV was proved to be a substrate of concentrative
nucleoside transporter (CNT) 2 and CNT3, of org. cation transporter (OCT) 3, and of breast cancer resistance protein
(BCRP) using transfected cells expressing resp. transporters. The inhibition of ETV uptake in primary human trophoblast
cells further confirmed that equilibrative nucleoside transporter (ENT) 1/2, CNT2/3, OCT3, and org. cation/carnitine
transporter (OCTN) 2 might be involved in ETV transfer in human placenta. Therefore, ETV uptake from maternal
circulation to trophoblast cells was possibly transported by CNT2/3, ENT1/2, and OCTN2, whereas ETV efflux from
trophoblast cells to fetal circulation was mediated by OCT3, and efflux from trophoblast cells to maternal circulation might
be mediated by BCRP, multidrug resistance-assocd. protein 2, and P-glycoprotein. The information obtained in the
present study may provide a basis for the use of ETV in pregnancy.
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9. Smurf1 regulates lung cancer cell growth and migration through interaction with and ubiquitination of PIPKIγ
By Li, H.; Xiao, N.; Wang, Y.; Wang, R.; Chen, Y.; Pan, W.; Liu, D.; Li, S.; Sun, J.; Zhang, K.; et al
From Oncogene (2017), Ahead of Print. Language: English, Database: CAPLUS, DOI:10.1038/onc.2017.166
Type Iγ phosphatidylinositol phosphate kinase (PIPKIγ), a phospholipid kinase generating PIP2, is pos. expressed in
breast cancer tissues, which correlates intimately with the progression of patients. However, little is known about the
expression level of PIPKIγ in patients with other cancer types as well as their underlying regulation mechanisms. Here,
we report that PIPKIγ is highly expressed in lung cancer tissues and its expression level is crit. for lung cancer cell
proliferation, which may serve as a prognostic marker for lung cancer patients. Meanwhile, we show that E3 ubiquitin
ligase Smurf1 directly interacts with PIPKIγ and targets PIPKIγ for ubiquitination and degrdn. in lung cancer cells. Also,
we discover that Smurf1 directly binds to the kinase domain of PIPKIγ via its C2 domain while Lysine 255 in PIPKIγ acts
as the major ubiquitin acceptor site for Smurf1. In addn., we demonstrate that the phosphorylation mimicking mutant of
Smurf1, Smurf1 T306D, prevents PIPKIγi2 from ubiquitination and subsequent degrdn. similar to the effect of forskolin-
potentiated cAMP formation, suggesting that Thr306 in Smurf1 is crit. for its phosphorylation by PKA. Moreover, PKA-
Smurf1-PIPKIγ signal transduction takes a significant part in lung cancer cell growth and in vivo tumorigenesis. Thus, we
propose that the PKA-Smurf1-PIPKIγ pathway has an important role in pulmonary tumorigenesis and imposes substantial
clin. impact on development of novel diagnostic markers and therapeutic targets for lung cancer treatment.
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10. Role of LKB1-CRTC1 on glycosylated COX-2 and response to COX-2 inhibition in lung cancer
By Cao, Chunxia; Gao, Ruli; Zhang, Min; Amelio, Antonio L.; Fallahi, Mohammad; Chen, Zirong; Gu, Yumei; Hu,
Chengbin; Welsh, Eric A.; Engel, Brienne E.; et al
From Journal of the National Cancer Institute (2015), 107(1), dju358/1-dju358/11. Language: English, Database:
CAPLUS, DOI:10.1093/jnci/dju358
Background: Cyclooxygenase-2 (COX-2) directs the synthesis of prostaglandins including PGE-2 linking inflammation
with mitogenic signaling. COX-2 is also an anticancer target, however, treatment strategies have been limited by
unreliable expression assays and by inconsistent tumor responses to COX-2 inhibition. Methods: We analyzed the
TCGA and Director's Challenge lung cancer datasets (n = 188) and also generated an LKB1-null lung cancer gene
signature (n = 53) to search the Broad Institute/Connectivity-MAP (C-MAP) dataset. We performed ChIP analyses, real-
time polymerase chain reaction, immunoblotting, and drug testing of tumor cell lines (n = 8) and primary lung
adenocarcinoma surgical resections (n = 13). Results: We show that COX-2 is a target of the cAMP/CREB coactivator
CRTC1 signaling pathway. In addn., we detected a correlation between LKB1 status, CRTC1 activation, and presence of
glycosylated, but not inactive hypoglycosylated COX-2 in primary lung adenocarcinoma. A search of the C-MAP drug
database discovered that all high-ranking drugs pos. assocd. with the LKB1-null signature are known CRTC1 activators,
including forskolin and six different PGE-2 analogs. Somatic LKB1 mutations are present in 20.0% of lung
adenocarcinomas, and we obsd. growth inhibition with COX-2 inhibitors in LKB1-null lung cancer cells with activated
CRTC1 as compared with LKB1-wildtype cells (NS-398, P = .002 and Niflumic acid, P = .006; two-tailed t test).
Conclusion: CRTC1 activation is a key event that drives the LKB1-null mRNA signature in lung cancer. We also
identified a pos. feedback LKB1/CRTC1 signaling loop for COX-2/PGE2 regulation. These data suggest a role for LKB1
status and glycosylated COX-2 as specific biomarkers that provide a framework for selecting patients for COX-2
inhibition studies.
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11. Type 2 inositol trisphosphate receptor gene expression in hepatocytes is regulated by cyclic AMP
By Kruglov, Emma; Ananthanarayanan, Meenakshisundaram; Sousa, Pedro; Weerachayaphorn, Jittima; Guerra,
Mateus T.; Nathanson, Michael H.
From Biochemical and Biophysical Research Communications (2017), 486(3), 659-664. Language: English, Database:
CAPLUS, DOI:10.1016/j.bbrc.2017.03.086
The type 2 inositol 1,4,5-trisphosphate receptor (IP3R2) is the principal intracellular Ca2+ release channel in hepatocytes,
and so is important for bile secretion and other functions. IP3R2 activity is regulated in part by post-translational
modifications but little is known about transcriptional regulation of its expression. We found that both IP3R2 mRNA and
protein levels in liver were increased during fasting. Treatment of hepatocytes with forskolin or 8-CPT-cAMP also
increased IP3R2, and this was reduced by actinomycin D. Anal. of the IP3R2 promoter revealed five CREs, and CREB
potently increased promoter activity. Mutation of CRE4 or CRE5 decreased induction by CREB, and ChIP assay showed
recruitment of CREB to these sites. Adenylyl cyclase (AC) 6 and 9 were the principal AC isoforms detected in rat
hepatocytes, and silencing either one decreased org. anion secretion, which depends on IP3R2. Secretion furthermore
was increased by overnight but not acute treatment with forskolin or 8-CPT-cAMP. These findings provide evidence that
IP3R2 expression is transcriptionally regulated by cAMP via CREB binding to CRE elements in its promoter. The
findings furthermore suggest that this mechanism is relevant for hormonal regulation of bile secretion.
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12. Three-dimensional hydrogels for culturing adult epithelial stem cells and organoids
By Gjorevski, Nikolche; Lutolf, Matthias
The invention provides a three-dimensional hydrogel for culturing adult epithelial stem cells and uses thereof.
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13. Prostaglandin E2-mediated adenosinergic effects on CD14+ cells: Self-amplifying immunosuppression in

cancer
By Al-Taei, Saly; Salimu, Josephine; Spary, Lisa K.; Clayton, Aled; Lester, Jason F.; Tabi, Zsuzsanna
From OncoImmunology (2017), 6(2), e1268308/1-e1268308/10. Language: English, Database: CAPLUS,
DOI:10.1080/2162402X.2016.1268308
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CD39 and CD73 are surface-expressed ectonucleotidases that hydrolyze ATP in a highly regulated, serial manner into
ADP, AMP and adenosine. The end product, adenosine, has both tumor-promoting and immunosuppressive effects.
The aim of this study was to det. CD73 expression on immune cells in pleural effusion (PE) in order to have a better
understanding of the immune environment in mesothelioma. PE- or blood-derived CD14+ cells of mesothelioma patients
and healthy donors were analyzed by flow cytometry for the expression of CD39 and CD73. CD73-induction was studied
by exposure of CD14+ cells to the sol. fraction of PE (sPE), while the signaling mechanism, responsible for CD73
induction, by phosphoflow cytometry and receptor-inhibition studies. We obsd. CD73 expression on CD14+ cells in PE
but not peripheral blood of mesothelioma patients or healthy donors. CD73 expression was inducible on CD14+ cells
with sPE, cyclic-AMP (cAMP)-inducers (forskolin and prostaglandin-E2 (PGE2)) and adenosine. Inhibition of PGE2
receptors or adenosine A2 receptors blocked CD73-induction by sPE. sPE treatment triggered protein kinase A and p38
activation. However, signal-transducer and activator of transcription 3 (STAT3)-blocking led to enhanced CD73
expression, demonstrating a hitherto unknown neg. control of purinergic signaling by STAT3 in CD14+ cells. TNFα
prodn. by CD73+ CD14+ cells was significantly impaired in the presence of AMP, confirming immunosuppressive
function. Taken together, CD73 expression can be induced by PGE2, cAMP or adenosine on human CD14+ cells. We
suggest that targeting this autocrine loop is a valid therapeutic approach in mesothelioma that may also enhance
immunotherapy.
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14. PKA/CREB regulates the constitutive promoter activity of the USP22 gene
By Xiong, Jianjun; Zhou, Xiaoou; Gong, Zhen; Wang, Ting; Zhang, Chao; Xu, Xiao Yuan; Liu, Jian Yun; Li, Weidong
From Oncology Reports (2015), 33(3), 1505-1511. Language: English, Database: CAPLUS, DOI:10.3892/or.2015.3740
The human ubiquitin-specific processing enzyme 22 (USP22) plays a crucial role in regulating cell cycle processes and
its overexpression has been linked to tumor progression. However, the mechanisms leading to USP22 transcriptional
activation in human cancer cells are still unclear. Previously, we characterized the 5'-flanking sequence of the human
USP22 gene and found a potential CREB/ATF binding site within the basic promoter region. The present study found
that this site was required for constitutive USP22 transcriptional activity in HeLa and HepG2 cells. Chromatin
immunopptn. assay confirmed that CREB interacted with this site. siRNA knockdown of CREB decreased USP22
transcriptional activation and endogenous expression, whereas CREB over-expression did not affect transcriptional
levels. Furthermore, USP22 promoter activity and expression were decreased by inhibiting PKA with H-89, but were not
responsive to forskolin induction. All of these results demonstrate that PKA/CREB is involved in the regulation of
constitutive promoter activity of the USP22 gene.
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15. Adenylyl cyclase 3/adenylyl cyclase-associated protein 1 (CAP1) complex mediates the anti-migratory effect
of forskolin in pancreatic cancer cells
By Quinn, Sierra N.; Graves, Sarai H.; Dains-McGahee, Clayton; Friedman, Emilee M.; Hassan, Humma; Witkowski,
Piotr; Sabbatini, Maria E.
From Molecular Carcinogenesis (2017), 56(4), 1344-1360. Language: English, Database: CAPLUS,
DOI:10.1002/mc.22598
Pancreatic cancer is one of the most lethal human malignancies. A better understanding of the intracellular mechanism
of migration and invasion is urgently needed to develop treatment that will suppress metastases and improve overall
survival. Cyclic adenosine monophosphate (cAMP) is a second messenger that has shown to regulate migration and
invasion of pancreatic cancer cells. The rise of cAMP suppressed migration and invasion of pancreatic ductal
adenocarcinoma cells. CAMP is formed from cytosolic ATP by the enzyme adenylyl cyclase (AC). There are ten
isoforms of ACs; nine are anchored in the plasma membrane and one is sol. What remains unknown is the extent to
which the expression of transmembrane AC isoforms is both modified in pancreatic cancer and mediates the inhibitory
effect of forskolin on cell motility. Using real-time PCR anal., ADCY3 was found to be highly expressed in pancreatic
tumor tissues, resulting in a constitutive increase in cAMP levels. On the other hand, ADCY2 was down-regulated.
Migration, invasion, and filopodia formation in two different pancreatic adenocarcinoma cell lines, HPAC and PANC-1
deficient in AC1 or AC3, were studied. We found that AC3, upon stimulation with forskolin, enhanced cAMP levels and
inhibited cell migration and invasion. Unlikely to be due to a cytotoxic effect, the inhibitory effects of forskolin involved the
quick formation of AC3/adenylyl cyclase-assocd. protein 1 (CAP1)/G-actin complex, which inhibited filopodia formation
and cell motility. Using Western blotting anal., forskolin, through AC3 activation, caused phosphorylation of CREB, but
not ERK. The effect of CREB phosphorylation is likely to be assocd. with long-term signaling changes. © 2016 Wiley
Periodicals, Inc.
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16. Pharmacological Activation of Protein Phosphatase 2 A (PP2A): A Novel Strategy to Fight Against Human
Malignancies?
By Carratu, Maria Rosaria; Signorile, Anna; De Rasmo, Domenico; Reale, Antonia; Vacca, Angelo
From Current Medicinal Chemistry (2016), 23(38), 4286-4296. Language: English, Database: CAPLUS,
DOI:10.2174/0929867323666161014133423
A review. Background: The serine-threonine protein phosphatase 2A (PP2A) regulates multiple cell signaling cascades
and its inactivation by viral oncoproteins, mutation of specific structural subunits or upregulation of the cellular
endogenous inhibitors may contribute to malignant transformation by regulating specific phosphorylation events.
Pharmacol. modulation of PP2A activity is becoming an attractive strategy for cancer treatment. Some compds. targeting
PP2A are able to induce PP2A reactivation and subsequent cell death in several types of cancer. Methods: We
undertook a search of bibliog. databases for peer-reviewed articles focusing on the main item of the review. We selected
articles published in indexed journals. The quality of retrieved papers was appraised using the std. bibliometric
indicators. Results: One hundred and fourteen papers were included in the review. Twenty-seven papers gave an
overview of structure and physiol. role of PP2A. Twenty-five papers outlined the role of PP2A in tumor suppression.
Forty papers analyzed the mechanism involved in PP2A reactivation by synthetic compds., and twenty-two papers
outlined the capability of natural compds. of restoring PP2A activity and how this could be beneficial. Conclusion:
Findings analyzed in this review underline the central role of PP2A as a regulator of cell growth and survival, hence its
function as tumor suppressor. The discovery that some compds., either synthetic or natural, are capable of reactivating
PP2A opens up new perspectives for future strategies to fully exploit therapeutic potential in human cancer. Thus, this
review could also be of particular interest to pharmaceutical or biotechnol. companies for drug design and targeted
delivery.
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17. The circadian clock modulates anti-cancer properties of curcumin

By Sarma, Ashapurna; Sharma, Vishal P.; Sarkar, Arindam B.; Sekar, M. Chandra; Samuel, Karunakar; Geusz,
Michael E.
From BMC Cancer (2016), 16, 759/1-759/13. Language: English, Database: CAPLUS, DOI:10.1186/s12885-016-2789-
9
Background: Curcuminoids of the spice turmeric and their enhanced derivs. have much potential as cancer treatments.
They act on a wide variety of biol. pathways, including those regulating cell division and circadian rhythms. It is known
that circadian clocks can modify cancer therapy effectiveness, according to studies aimed at optimizing treatments based
on the circadian cycle. It is therefore important to det. whether treatments with curcumin or similar chemotherapeutic
agents are regulated by circadian timing. Similarly, it is important to characterize any effects of curcumin on timing
abilities of the circadian clocks within cancer cells. Methods: We examd. the circadian clock's impact on the timing of cell
death and cell division in curcumintreated C6 rat glioma cells through continuous video microscopy for several days. To
evaluate its persistence and distribution in cancer cells, curcumin was localized within cell compartments by imaging its
autofluorescence. Finally, HPLC and spectroscopy were used to det. the relative stabilities of the curcumin congeners
demethoxycurcumin and bisdemethoxycurcumin that are present in turmeric. Results: Circadian rhythms in cell death
were obsd. in response to low (5 µM) curcumin, reaching a peak several hours before the peak in rhythmic expression of
mPER2 protein, a major circadian clock component. These results revealed a sensitive phase of the circadian cycle that
could be effectively targeted in patient therapies based on curcumin or its analogs. Curcumin fluorescence was obsd. in
cell compartments at least 24 h after treatment, and the two congeners displayed greater stability than curcumin in cell
culture medium. Conclusions: We propose a mechanism whereby curcuminoids act in a sustained manner, over several
days, despite their tendency to degrade rapidly in blood and other aq. media. During cancer therapy, curcumin or its
analogs should be delivered to tumor cells at the optimal phase for highest efficacy after identifying the circadian phase
of the cancer cells. We confirmed the greater stability of the curcumin congeners, suggesting that they may produce
sustained toxicity in cancer cells and should be considered for use in patient care.
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18. Activation of GPR30 stimulates GTP-binding of Gαi1 protein to sustain activation of Erk1/2 in inhibition of
prostate cancer cell growth and modulates metastatic properties
By Lau, Kin-Mang; Ma, Fanny Man-Ting; Xia, Jenny Tian; Chan, Queeny Kwan Yi; Ng, Chi-Fai; To, Ka-Fai
DOI:10.1016/j.yexcr.2016.11.022
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Previously, we reported that GPR30 activation by the receptor-specific, non-estrogenic ligand G-1 inhibited in vitro and in
vivo growth of prostate cancer (PCa) cells via sustained Erk1/2 activation. Mechanism underlying the sustained Erk1/2
activation for PCa cell growth inhibition remains unclear. Here we report that G-1, through GPR30, activated Gαi1
proteins to sustain Erk1/2 activation but failed to activate adenylyl cyclase (AC) for cAMP prodn. in PCa cells. The
chem.-induced activation of AC-cAMP-PKA signaling attenuated Erk1/2 activity and blocked the cell growth inhibitory
effects of G-1. Furthermore, PCa predominantly expressed Gαi1 proteins. Silencing of Gαi1 expression blocked the
inhibitory effects of G-1 on PCa cell growth. By gene expression profiling, GPR30 activation by G-1 interfered expression
of cell cycle regulators and machinery elements to modulate PCa cell growth and the RACGAP1 interactome to control
metastatic properties. In this regard, we demonstrated that G-1 inhibited PCa cell migration and invasion with reduced
formations of filopodia and stress fibers through a GPR30-dependent pathway. Taken together, our findings revealed the
underlying mechanism for sustaining Erk1/2 activation upon GPR30 activation by G-1 in PCa cells and the GPR30-
mediated pathways in controlling PCa cell growth and metastatic properties.
~1 Citing
19. Characterization of Hippo Pathway Components by Gene Inactivation

By Plouffe, Steven W.; Meng, Zhipeng; Lin, Kimberly C.; Lin, Brian; Hong, Audrey W.; Chun, Justin V.; Guan, Kun-
Liang
From Molecular Cell (2016), 64(5), 993-1008. Language: English, Database: CAPLUS,
DOI:10.1016/j.molcel.2016.10.034
The Hippo pathway is important for regulating tissue homeostasis, and its dysregulation has been implicated in human
cancer. However, it is not well understood how the Hippo pathway becomes dysregulated because few mutations in core
Hippo pathway components have been identified. Therefore, much work in the Hippo field has focused on identifying
upstream regulators, and a complex Hippo interactome has been identified. Nevertheless, it is not always clear which
components are the most physiol. relevant in regulating YAP/TAZ. To provide an overview of important Hippo pathway
components, we created knockout cell lines for many of these components and compared their relative contributions to
YAP/TAZ regulation in response to a wide range of physiol. signals. By this approach, we provide an overview of the
functional importance of many Hippo pathway components and demonstrate NF2 and RHOA as important regulators of
YAP/TAZ and TAOK1/3 as direct kinases for LATS1/2.
~4 Citings
20. CXC chemokine receptor 3 alternative splice variants selectively activate different signaling pathways
By Berchiche, Yamina A.; Sakmar, Thomas P.
From Molecular Pharmacology (2016), 90(4), 483-495. Language: English, Database: CAPLUS,
DOI:10.1124/mol.116.105502
The G protein-coupled receptor (GPCR) C-X-C chemokine receptor 3 (CXCR3) is a potential drug target that mediates
signaling involved in cancer metastasis and inflammatory diseases. The CXCR3 primary transcript has three potential
alternative splice variants and cell-type specific expression results in receptor variants that are believed to have different
functional characteristics. However, the mol. pharmacol. of ligand binding to CXCR3 alternative splice variants and their
downstream signaling pathways remain poorly explored. To better understand the role of the functional consequences of
alternative splicing of CXCR3, we measured signaling in response to four different chemokine ligands (CXCL4, CXCL9,
CXCL10, and CXCL11) with agonist activity at CXCR3. Both CXCL10 and CXCL11 activated splice variant CXCR3A.
Whereas CXCL10 displayed full agonistic activity for Gαi activation and extracellular signal regulated kinase (ERK) 1/2
phosphorylation and partial agonist activity for β-arrestin recruitment, CXCL9 triggered only modest ERK1/2
phosphorylation. CXCL11 induced CXCR3B-mediated β-arrestin recruitment and little ERK phosphorylation. CXCR3Alt
signaling was limited to modest ligand-induced receptor internalization and ERK1/2 phosphorylation in response to
chemokines CXCL11, CXCL10, and CXCL9. These results show that CXCR3 splice variants activate different signaling
pathways and that CXCR3 variant function is not redundant, suggesting a mechanism for tissue specific biased agonism.
Our data show an addnl. layer of complexity for chemokine receptor signaling that might be exploited to target specific
CXCR3 splice variants.
~2 Citings
21. Increase of Intracellular Cyclic AMP by PDE4 Inhibitors Affects HepG2 Cell Cycle Progression and Survival
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By Massimi, Mara; Cardarelli, Silvia; Galli, Francesca; Giardi, Maria Federica; Ragusa, Federica; Panera, Nadia;
Cinque, Benedetta; Cifone, Maria Grazia; Biagioni, Stefano; Giorgi, Mauro
From Journal of Cellular Biochemistry (2017), 118(6), 1401-1411. Language: English, Database: CAPLUS,
DOI:10.1002/jcb.25798
Type 4 cyclic nucleotide phosphodiesterases (PDE4) are major members of a superfamily of enzymes (PDE) involved in
modulation of intracellular signaling mediated by cAMP. Broadly expressed in most human tissues and present in large
amts. in the liver, PDEs have in the last decade been key therapeutic targets for several inflammatory diseases.
Recently, a significant body of work has underscored their involvement in different kinds of cancer, but with no attention
paid to liver cancer. The present study investigated the effects of two PDE4 inhibitors, rolipram and DC-TA-46, on the
growth of human hepatoma HepG2 cells. Treatment with these inhibitors caused a marked increase of intracellular
cAMP level and a dose- and time-dependent effect on cell growth. The concns. of inhibitors that halved cell proliferation
to about 50% were used for cell cycle expts. Rolipram (10 µM) and DC-TA-46 (0.5 µM) produced a decrease of cyclin
expression, in particular of cyclin A, as well as an increase in p21, p27 and p53, as evaluated by Western blot anal.
Changes in the intracellular localization of cyclin D1 were also obsd. after treatments. In addn., both inhibitors caused
apoptosis, as demonstrated by an Annexin-V cytofluorimetric assay and anal. of caspase-3/7 activity. Results
demonstrated that treatment with PDE4 inhibitors affected HepG2 cell cycle and survival, suggesting that they might be
useful as potential adjuvant, chemotherapeutic or chemopreventive agents in hepatocellular carcinoma. J. Cell.
Biochem. 9999: 1-11, 2016. © 2016 Wiley Periodicals, Inc.
~1 Citing
22. Label free biosensors and cells

By Fang, Ye; Ferrie, Ann M.; Fontaine, Norman H.; Lahiri, Joydeep; Yuen, Po Ki
From U.S. (2011), US 8076090 B2 20111213, Language: English, Database: CAPLUS
Disclosed are compns. and methods for using label free optical biosensors for performing cell assays. In certain
embodiments the assays can be performed in high throughput methods and can be multiplexed.
~4 Citings
23. The Natural cAMP Elevating Compound Forskolin in Cancer Therapy: Is It Time?
By Sapio, Luigi; Gallo, Monica; Illiano, Michela; Chiosi, Emilio; Naviglio, Daniele; Spina, Annamaria; Naviglio, Silvio
From Journal of Cellular Physiology (2017), 232(5), 922-927. Language: English, Database: CAPLUS,
DOI:10.1002/jcp.25650
A review. Cancer is a major public health problem and the second leading cause of mortality around the world. Although
continuous advances in the science of oncol. and cancer research are now leading to improved outcomes for many
cancer patients, novel cancer treatment options are strongly demanded. Naturally occurring compds. from a variety of
vegetables, fruits, and medicinal plants have been shown to exhibit various anticancer properties in a no. of in vitro and
in vivo studies and represent an attractive research area for the development of new therapeutic strategies to fight
cancer. Forskolin is a diterpene produced by the roots of the Indian plant Coleus forskohlii. The natural compd. forskolin
has been used for centuries in traditional medicine and its safety has also been documented in conventional modern
medicine. Forskolin directly activates the adenylate cyclase enzyme, that generates cAMP from ATP, thus, raising
intracellular cAMP levels. Notably, cAMP signaling, through the PKA-dependent and/or -independent pathways, is very
relevant to cancer and its targeting has shown a no. of antitumor effects, including the induction of mesenchymal-to-
epithelial transition, inhibition of cell growth and migration and enhancement of sensitivity to conventional antitumor drugs
in cancer cells. Here, we describe some features of cAMP signaling that are relevant to cancer biol. and address the
state of the art concerning the natural cAMP elevating compd. forskolin and its perspectives as an effective anticancer
agent. J. Cell. Physiol. 9999: 1-6, 2016. © 2016 Wiley Periodicals, Inc.
~2 Citings
24. Identification of phosphorylation sites regulating sst3 somatostatin receptor trafficking

By Lehmann, Andreas; Kliewer, Andrea; Guenther, Thomas; Nagel, Falko; Schulz, Stefan
From Molecular Endocrinology (2016), 30(6), 645-659. Language: English, Database: CAPLUS,
DOI:10.1210/me.2015-1244
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The human somatostatin receptor 3 (sst3) is expressed in about 50% of all neuroendocrine tumors and hence a
promising target for multireceptor somatostatin analogs. The sst3 receptor is unique among ssts in that it exhibits a very
long intracellular C-terminal tail contg. a huge no. of potential phosphate acceptor sites. Consequently, our knowledge
about the functional role of the C-terminal tail in sst3 receptor regulation is very limited. Here, we have generated a
series of phosphorylation-deficient mutants that enabled us to det. crucial sites for its agonistinduced _-arrestin
mobilization, internalization, and down-regulation. Based on this information, we generated phosphosite-specific
antibodies for C-terminal Ser337/Thr341, Thr348, and Ser361 that enabled us to investigate the temporal patterns of sst3
phosphorylation and dephosphorylation. We found that the endogenous ligand somatostatin induced a rapid and robust
phosphorylation that was completely blocked by the sst3 antagonist NVP-ACQ090. The stable somatostatin analogs
pasireotide and octreotide promoted clearly less phosphorylation compared with somatostatin. We also show that sst3
phosphorylation occurred within seconds to minutes, whereas dephosphorylation of the sst3 receptor occurred at a
considerable slower rate. In addn., we also identified G protein-coupled receptor kinases 2 and 3 and protein
phosphatase 1α and 1β as key regulators of sst3 phosphorylation and dephosphorylation, resp. Thus, we here define
the C-terminal phosphorylation motif of the human sst3 receptor that regulates its agonist-promoted phosphorylation, β-
arrestin recruitment, and internalization of this clin. relevant receptor.
~2 Citings
25. Patient treatment via teratogenic pharmaceutical compounds or stem cell differentiating agents to induce
destructive pathways for treatment of disease stem cells
By Soon-Shiong, Patrick
Compns. and methods for treatment of a condition assocd. with disease stem cells, and esp. cancer stem cells are
disclosed. In one aspect, a patient is treated with a stem cell differentiating agent and/or teratogenic pharmaceutical
compd. to induce one or more destructive pathways in the disease stem cells. Most typically, the destructive pathways
include apoptotic pathways, necrotic pathways, and autophagy pathways.
~0 Citings
26. Methods for extending lifespan and methods of screening known pharmacological agents for new uses
By Ryazanov, Alexey G.; Chikunov, Alexander V.; Shymkiv, Yuriy
The present disclosure relates to methods of extending the
lifespan of a subject or treating, suppressing, inhibiting or
delaying the onset of an age-related condition or disorder, such
as cancer, in a subject. The methods comprise administering
to the subject an effective amt. of (i) a compd. that sustains
pharmacol. activation of xenobiotic metab., (ii) a cardiac
glycoside, (iii) a chelator, (iv) inulin, (v) D-valine, or any
combination thereof. In a further aspect, the present disclosure
relates to a method of identifying clin. candidates by performing
a high throughput screening in mammals of test compds. which
have great structural and/or functional diversity.
~0 Citings
27. Effects of tris(2,3-dibromopropyl) isocyanurate on steroidogenesis in H295R cells

By Li, Xun; Pan, Yu; Wang, Chang; Chen, Minjie; Liu, Yuchen; Li, Jia; Zhou, Zhen; Xu, Jinhua; Liang, Yong; Song,
Maoyong
From Environmental Earth Sciences (2016), 75(20), 1-8. Language: English, Database: CAPLUS,
DOI:10.1007/s12665-016-6166-4
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Tris(2,3-dibromopropyl) isocyanurate (TBC) is widely used in polymer products. It is ubiquitously found in environmental
matrixes. Previous studies have shown that TBC has potential endocrine-disrupting effects on aquatic organisms.
However, the effects on adrenocortical function and the underlying mechanisms are not well characterized. In present
study, the H295R cell line was employed to investigate the potential endocrine-disrupting action of TBC. TBC inhibited
basal and forskolin-induced sex hormone prodn. but did not exhibit cytotoxicity. The expression of the steroidogenic
genes, StAR, 3βHSD2, CYP17, CYP21, CYP19, HMGR, 17βHSD1, 17βHSD4 and CYP11A, was not changed upon TBC
exposure. However, the cAMP inducer forskolin strongly induced the expression of all these genes, except for HMGR,
17βHSD1 and 17βHSD4. TBC blocked forskolin-induced StAR, 3βHSD2, CYP17, CYP21 and CYP19 mRNA
expression. Our results indicate that TBC can inhibit sex hormone biosynthesis due to the interference with
steroidogenic gene transcription in activated condition.
~0 Citings
28. Genomic analyses identify agents regulating somatotroph and lactotroph functions
By Fan, Jun; Zhang, Cui; Chen, Qi; Zhou, Jin; Franc, Jean-Louis; Chen, Qing; Tong, Yunguang
From Functional & Integrative Genomics (2016), 16(6), 693-704. Language: English, Database: CAPLUS,
DOI:10.1007/s10142-016-0518-8
Isolated hormone deficiency might be caused by loss of a specific type of endocrine cells, and regenerating these
missing cells may provide a new option for future treatment. It is known that POU1F1 lineage cells can differentiate into
thyrotroph, somatotroph, and lactotroph. However, there is no effective way of controlling pituitary stem/progenitor cells
to differentiate into a specific type of endocrine cell. We thereby analyzed multiple genomic publications related to
POU1F1 and pituitary development in this study to identify genes and agents regulating POU1F1 lineage cell
differentiation. ANOVA analyses were performed to obtain differentially expressed genes. Ingenuity pathway analyses
were performed to obtain signaling pathways, interaction networks, and upstream regulators. Venn diagram was used to
det. the overlapping information between studies. Summary statistics was performed to rank genes according to their
frequency of occurrence in these studies. The results from upstream analyses indicated that 326 agents may regulate
pituitary cell differentiation. These agents can be categorized into 12 groups, including hormones and related pathways,
PKA-cAMP pathways, p53/DNA damaging/cell cycle pathways, immune/inflammation regulators, growth factor and
downstream pathways, retinoic/RAR pathways, ROS pathways, histone modifications, CCAAT/enhancer binding protein
family, neuron development/degeneration pathways, calcium related and fat acid, and glucose pathways. Addnl. expts.
demonstrated that H2O2 and catalase differentially regulate growth hormone and prolactin expression in
somatolactotroph cells, confirming potential roles of ROS pathway on regulating somatotroph and lactotroph functions.
~0 Citings
29. In situ PKA activity assay by selective detection of its catalytic subunit using antibody arrays
By Sayyed, Danishmalik Rafiq; Jung, Se-Hui; Kim, Min-Soo; Han, Eun-Taek; Park, Won Sun; Hong, Seok-Ho; Kim,
Young-Myeong; Ha, Kwon-Soo
From BioChip Journal (2017), 11(1), 57-66. Language: English, Database: CAPLUS, DOI:10.1007/s13206-016-1108-5
Protein kinase A (PKA) plays a pivotal role in various biol. processes and the pathogenesis of several diseases.
However, systematic investigation of PKA functions in cells and tissues is limited due to the lack of a suitable high-
throughput in situ PKA activity assay. Here, we present an array-based in situ PKA activity assay that employs selective
detection of the catalytic form of PKA (cPKA; active and autophosphorylated) using antibody arrays. Antibody arrays
were fabricated by applying anti-cPKA antibody onto welltype amine arrays. The limit of detection was 0.02 µg/ mL. We
successfully applied this assay to det. changes in intracellular and extracellular PKA activities in human gastric
adenocarcinoma (AGS) cells and human umbilical vein endothelial cells (HUVECs) treated with forskolin. Forskolin
induced activation of intracellular PKA in a dose-dependent manner, and this PKA activation was inhibited by the potent
PKA inhibitor H-89. Extracellular PKA activity was also elevated by forskolin in a dose-dependent manner in AGS cells,
but this elevation was hardly detectable in HUVECs. Thus, our antibody array-based in situ PKA activity assay is
suitable for investigating the regulation and functions of intracellular and extracellular PKA in cells and tissues and has a
potential for use in cancer diagnosis.
~0 Citings
30. Repurposing treprostinil for enhancing hematopoietic progenitor cell transplantation

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By Kazemi, Zahra; Bergmayr, Christian; Prchal-Murphy, Michaela; Javaheri, Tahereh; Themanns, Madeleine; Pham,
Ha T. T.; Strohmaier, Wolfgang; Sexl, Veronika; Freissmuth, Michael; Zebedin-Brandl, Eva
DOI:10.1124/mol.116.103267
Activation of Gs-coupled receptors enhances engraftment of hematopoietic stem and progenitor cells (HSPCs). We
tested the hypothesis that treprostinil, a prostacyclin analog approved for the treatment of pulmonary hypertension, can
be repurposed to improve hematopoietic stem cell transplantation. Murine and human HSPCs were isolated from bone
marrow and umbilical cord blood, resp. Prostanoid receptor agonists and the combination thereof with forskolin were
tested for their capacity to stimulate [3H]cAMP accumulation in HSPCs. Three independent approaches were employed
to verify the ability of agonist-activated HSPCs to reconstitute the bone marrow in lethally irradiated recipient mice. The
underlyingmechanism was explored in cellularmigration assays and by blocking C-X-C motif chemokine receptor 4
(CXCR4). Among several prostanoid agonists tested in combination with forskolin, treprostinil was most efficacious in
raising intracellular cAMP levels in murine and human HPSCs. Injection of murine and human HSPCs, which had been
pretreated with treprostinil and forskolin, enhanced survival of lethally irradiated recipient mice. Survival was further
improved if recipientmice were s.c. administered treprostinil (0.15 mg kg-1 8 h-1) for 10 days. This regimen also reduced
the no. of HSPCs required to rescue lethally irradiated mice. Enhanced survival of recipient mice was causally related to
treprostinil-enhanced CXCR4-dependent migration of HSPCs. Treprostinil stimulates the engraftment of human and
murine hematopoietic stem cells without impairing their capacity for self-renewal. The investigated dose range
corresponds to the dose approved for human use. Hence, these findings may be readily translated into a clin.
application.
~0 Citings
31. Dye-stabilized nanoparticles and methods of their manufacture and therapeutic use
By Heller, Daniel A.; Shamay, Yosef
Described herein are nanoparticles which are largely made of
(e.g., 90 wt%) hydrophobic drugs and are stabilized by water
sol. dyes. The nanoparticles can range in size from 30 nm to
150 nm and have highly neg. surface charge (e.g., -55 mV).
These nanoparticles are highly sol. in water, stable for days in
PBS buffer and can be easily lyophilized and reconstituted in
water. Using quant. self-assembly prediction calcns.,
topochem. mol. descriptors were identified and validated as
highly predictive indicators of nano-assembly, nanoparticle
size, and drug loading. The resulting nanoparticles selectively
targeted kinase inhibitors to caveolin-1-expressing human
colon cancer and autochthonous liver cancer models to yield
striking therapeutic effects while avoiding pERK inhibition in
healthy skin. The nanoparticles exhibited remarkable anti-
tumor efficacy in vitro and in vivo in models of hepatocellular
carcinoma. Thus, dye-encapsulated sorafenib nanoparticles
(IR783-SFB) were synthesized using the nanopptn. method;
0.1 mL of sorafenib dissolved in DMSO (10 mg/mL) was added
dropwise (20 µL per 15 s) to a 0.4 aq. IR783 soln. (1 mg/mL)
contg. 0.05 mM sodium bicarbonate; the soln. was centrifuged
twice and resuspension in 1 mL of sterile PBS; the suspension
of nanoparticles was sonicated for 10 s with a probe sonicator
at 40 % intensity; the resulted IR783-SFB nanoparticles had
zeta potential of -52 mV and a size of 95 nm with a PDI of 0.1;
by suspending the nanoparticles in lower vols. they were able
to solubilize sorafenib up to 16 mg/mL in saline soln. which is
2000 times better than free drug; the nanoparticles were easily
lyophilized with a saline/sucrose 5 % soln. and reconstituted in
water at this concn.
~0 Citings
32. Phthalazinone pyrazole derivatives for treating retinal degenerative disease

By Zack, Donald J.; Fuller, John A.
SciFinder® Page 14
Compds., compns., kits and methods for treating conditions related to Retinal degenerations, and related diseases,
including retinitis pigmentosa and atrophic age-related macular degeneration, are disclosed.
~0 Citings
33. Piperazine-based alpha-1 AR blocker, naftopidil, selectively suppresses malignant human bladder cells via
induction of apoptosis
By Nakagawa, Yusuke U.; Nagaya, Hisao; Miyata, Takeaki; Wada, Yoshitaka; Oyama, Tsunehiro; Gotoh, Akinobu
From Anticancer Research (2016), 36(4), 1563-1570. Language: English, Database: CAPLUS
A retrospective observational cohort study has shown that exposure to alpha-1 adrenergic receptor (AR) blocker reduces
the risk of bladder cancer (BCa). We investigated the antitumor activity of alpha-1 blockers, that are administered long-
term therapeutically, in BCa. The antitumor activity of alpha-1 blockers was evaluated in terms of cell viability, cell cycle,
competition, and apoptotic signaling in BCa cells. Our cell viability studies showed that naftopidil was one of the
strongest alpha-1 AR blockers, regarding its antitumor action in BCa cells, independent of the grade of malignancy, but
with no similar action on normal human bladder cells. Oral administration of naftopidil reduced tumor vol. in a xenograft
model. Our own competitive anal. using an alpha-1 AR agonist and other alpha-1 AR blockers showed that naftopidil
activated cell death signaling without inhibitory action on alpha-1 ARs. We conclude that naftopidil has potential as an
antitumor drug against BCa in vitro and in vivo. This finding provides a rationale for developing naftopidil in grade-
independent treatment of BCa.
~0 Citings
34. Gingival epithelial cells support osteoclastogenesis by producing receptor activator of nuclear factor kappa
B ligand via protein kinase A signaling
By Usui, M.; Sato, T.; Yamamoto, G.; Okamatsu, Y.; Hanatani, T.; Moritani, Y.; Sano, K.; Yamamoto, M.; Nakashima,
K.
From Journal of Periodontal Research (2016), 51(4), 462-470. Language: English, Database: CAPLUS,
DOI:10.1111/jre.12323
Background and Objective : Periodontal disease is dental plaque-induced inflammatory disease of the periodontal tissues
that results in bone loss in the affected teeth. During bone resorption, receptor activator of nuclear factor kappa B ligand
(RANKL) is an essential factor that regulates osteoclastogenesis. Recently, we found that gingival epithelial cells (GECs)
in periodontal tissue produce RANKL, the expression of which is regulated by tumor necrosis factor-α and protein kinase
A signaling. In this study, we asked whether RANKL-producing GECs induce bone marrow macrophages (BMMs) to
form osteoclasts in a co-culture system. Material and Methods : Ca9-22 GECs and osteoclast precursor BMMs were co-
cultured with or without the protein kinase A signaling activator forskolin or inhibitor H89 to examine whether the RANKL-
producing GECs could be induced to form osteoclasts, as detd. using a pit formation assay. Results : Osteoclasts
formed spontaneously in co-cultures of Ca9-22 cells and BMMs, even in the absence of RANKL. The cells were cultured
on bone slices for 14 d, at which time resorption pits were obsd. Forskolin treatment significantly increased osteoclast
nos. in these co-cultures, but forskolin alone did not induce osteoclast formation by BMMs. Conclusion : GECs
producing RANKL are able to support osteoclastogenesis in an in vitro co-culture system using GECs and BMMs, in a
process promoted by forskolin.
~1 Citing
35. High-throughput screening of chemical effects on steroidogenesis using H295R human adrenocortical
carcinoma cells
By Karmaus, Agnes L.; Toole, Colleen M.; Filer, Dayne L.; Lewis, Kenneth C.; Martin, Matthew T.
From Toxicological Sciences (2016), 150(2), 323-332. Language: English, Database: CAPLUS,
DOI:10.1093/toxsci/kfw002
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Disruption of steroidogenesis by environmental chems. can result in altered hormone levels causing adverse
reproductive and developmental effects. A high-throughput assay using H295R human adrenocortical carcinoma cells
was used to evaluate the effect of 2060 chem. samples on steroidogenesis via high-performance liq. chromatog. followed
by tandem mass spectrometry quantification of 10 steroid hormones, including progestagens, glucocorticoids,
androgens, and estrogens. The study employed a 3 stage screening strategy. The first stage established the max.
tolerated concn. (MTC; ≥ 70% viability) per sample. The second stage quantified changes in hormone levels at the MTC
whereas the third stage performed concn.-response (CR) on a subset of samples. At all stages, cells were prestimulated
with 10 µM forskolin for 48h to induce steroidogenesis followed by chem. treatment for 48 h. Of the 2060 chem. samples
evaluated, 524 samples were selected for 6-point CR screening, based in part on significantly altering at least 4
hormones at the MTC. CR screening identified 232 chem. samples with concn.-dependent effects on 17β-estradiol
and/or testosterone, with 411 chem. samples showing an effect on at least one hormone across the steroidogenesis
pathway. Clustering of the concn.-dependent chem.-mediated steroid hormone effects grouped chem. samples into 5
distinct profiles generally representing putative mechanisms of action, including CYP17A1 and HSD3B inhibition. A
distinct pattern was obsd. between imidazole and triazole fungicides suggesting potentially distinct mechanisms of action.
From a chem. testing and prioritization perspective, this assay platform provides a robust model for high-throughput
screening of chems. for effects on steroidogenesis.
~5 Citings
36. P-Hydroxycinnamaldehyde Induces B16-F1 Melanoma Cell Differentiation via the RhoA-MAPK Signaling
Pathway
By Zhao, Lian-mei; Sun, Guo-gui; Han, Li-na; Liu, Li-hua; Ren, Feng-zhi; Li, Lei; Ma, Ming; Shan, Bao-en
From Cellular Physiology and Biochemistry (2016), 38(6), 2247-2260. Language: English, Database: CAPLUS,
DOI:10.1159/000445580
Background/Aims: Due to its antitumor and gastroprotective properties, cochinchina momordica seed (CMS), has been
widely used to treat cancer patients in Asia. Our previous reports have shown that CMS is able to induce the
differentiation of B16-F1 melanoma cells. However, its functional component and mechanism remain unclear and are
addressed in this study. Methods and Results: CMSP (p-hydroxycinnamaldehyde isolated from CMS) inhibited the
proliferation, migration and invasiveness of B16-F1 cells both in vivo and in vitro. CMSP also induced the differentiation
of B16-F1 cells, as characterized by dendrite-like outgrowth, increased melanogenesis and enhanced tyrosinase activity.
Furthermore, CMSP treatment reduced the level of malignant markers of melanoma, specifically S-100B and melanoma-
derived growth regulatory protein precursor (MIA), in a concn.-dependent manner. According to a western blot anal.,
B16-F1 cells treated with CMSP exhibited a sustained increase in p-P38 and decreased activities of ERK and JNK. Our
data further indicated that the downregulation of GTP-RhoA, which was mediated by increased cAMP release, was
involved in CMSP-induced changes in MAPK, while LPA (Lysophosphatidic acid) partially reversed CMSP-induced B16
cell differentiation. Conclusion: These results demonstrated that CMSP-induced differentiation of B16F1 cells may occur
through the RhoA-MAPK axis, which suggests a new potential strategy for melanoma treatment.
~1 Citing
37. A CIEF-LIF method for simultaneous analysis of multiple protein kinases and screening of inhibitors
By Xiao, Lixia; Liu, Shengquan; Lin, Lihua; Yao, Shouzhuo
From Electrophoresis (2016), 37(14), 2075-2082. Language: English, Database: CAPLUS,
DOI:10.1002/elps.201600090
Here, a CIEF-LIF method for multiple protein kinase simultaneous anal. and inhibitors throughput screening with fast rate
and low cost is presented. Comparing with CZE, CIEF-LIF exhibited great focusing ability and high sepn. efficiency for
substrate and phosphorylated peptides, and is applicable for multiple kinases simultaneous anal. regardless of their
substrate peptides compns. and charge statuses. Thus, highly sensitive anal. for cyclic adenosine 3', 5'-monophosphate-
dependent protein kinase (PKA) and cyclin-dependent kinase 1 (CDK1) was achieved in CIEF-LIF anal. with detection
sensitivity up to 1.25 mU/µL and 0.4 mU/µL, resp., two magnitudes higher than that of CZE and comparable with that in
nanomaterials or green fluorescent protein-based kinase assay. Moreover, the inhibition effect of inhibitors on multiple
kinases could be simultaneously readout in a single electrophoretic run, with half maximal inhibitory concn. of H-89 for
PKA and Ro-3306 for CDK1 calcd. as 37.0 and 35.9 nM, resp., consistent with literatures reported. The CIEF-LIF also
exhibited strong anti-interference ability in human breast cancer cell lysates anal. and simulators such as forskolin and 3-
isobutyl-1-methylxantine assessment. Therefore, CIEF-LIF is desirable for future biol. application and clin. diagnostics
and drug discovery.
~0 Citings

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38. Method for treating a brain tumour using dopamine D4 receptor (DRD4) antagonist in combination other
chemotherapeutic agents
By Dirks, Peter; Dolma, Sonam
A method of treating a brain tumor such as glioblastoma in a
mammal is provided comprising administering to the mammal a
dopamine D4 receptor (DRD4) antagonist (e.g. PNU-96415E,
fananserin, L-741742).
~0 Citings
39. Method for inhibiting survival, tumorigenesis and metastasis of cancer cells and/or cancer stem cells
By Hung, Shih-Chieh
The present invention provides a new method for inhibiting and preventing survival, tumorigenesis and metastasis of
cancer stem cells (CSCs) in a subject using Collagen XVII (Col XVII) inhibitor to CSCs, or a use of Col XVII inhibitor for
the manuf. of a medicament for inhibiting and preventing survival, tumorigenesis and metastasis of CSCs. The present
invention also provides a pharmaceutical compn. for inhibiting and preventing survival, tumorigenesis and metastasis of
cancer stem cells, comprising an effective amt. of Col XVII inhibitor.
~0 Citings
40. Label-Free Functional Selectivity Assays

By Ferrie, Ann M.; Goral, Vasiliy; Wang, Chaoming; Fang, Ye
From Methods in Molecular Biology (New York, NY, United States) (2015), 1272(G Protein-Coupled Receptor
Screening Assays), 227-246. Language: English, Database: CAPLUS, DOI:10.1007/978-1-4939-2336-6_16
G protein-coupled receptors (GPCRs) represent the largest class of drug targets. Ligand-directed functional selectivity or
biased agonism opens new possibility for discovering GPCR drugs with better efficacy and safety profiles. However,
quantification of ligand bias is challenging. Herein, we present five different label-free dynamic mass redistribution
(DMR) approaches to assess ligand bias acting at the β2 -adrenergic receptor (β2 AR). Multiparametric anal. of the DMR
agonist profiles reveals divergent pharmacol. of a panel of β2 AR agonists. DMR profiling using catechol as a
conformational probe detects the presence of multiple conformations of the β2 AR. DMR assays under microfluidics,
together with chem. biol. tools, discover ligand-directed desensitization of the receptor. DMR antagonist reverse assays
manifest biased antagonism. DMR profiling using distinct probe-modulated cells detects the biased agonism in the
context of self-referenced pharmacol. activity map.
~4 Citings
41. Adrenergic Stimulation of DUSP1 Impairs Chemotherapy Response in Ovarian Cancer

By Kang, Yu; Nagaraja, Archana S.; Armaiz-Pena, Guillermo N.; Dorniak, Piotr L.; Hu, Wei; Rupaimoole, Rajesha; Liu,
Tao; Gharpure, Kshipra M.; Previs, Rebecca A.; Hansen, Jean M.; et al
From Clinical Cancer Research (2016), 22(7), 1713-1724. Language: English, Database: CAPLUS, DOI:10.1158/1078-
0432.CCR-15-1275
SciFinder® Page 17
Purpose: Chronic adrenergic activation has been shown to assoc. with adverse clin. outcomes in cancer patients, but the
underlying mechanisms are not well understood. The focus of the current study was to det. the functional and biol.
effects of adrenergic pathways on response to chemotherapy in the context of ovarian cancer. Exptl. Design: Increased
DUSP1 prodn. by sympathetic nervous system mediators (e.g., norepinephrine) was analyzed by real-time quant. RT-
PCR and by Western blotting. In vitro chemotherapy-induced cell apoptosis was examd. by flow cytometry. For in vivo
therapy, a well-characterized model of chronic stress was used. Results: Catecholamines significantly inhibited
paclitaxel- and cisplatin-induced apoptosis in ovarian cancer cells. Genomic analyses of cells treated with
norepinephrine identified DUSP1 as a potential mediator. DUSP1 overexpression resulted in reduced paclitaxel-induced
apoptosis in ovarian cancer cells compared with control; conversely, DUSP1 gene silencing resulted in increased
apoptosis compared with control cells. DUSP1 gene silencing in vivo significantly enhanced response to paclitaxel and
increased apoptosis. In vitro analyses indicated that norepinephrine-induced DUSP1 gene expression was mediated
through ADRB2 activation of cAMP-PLC-PKC-CREB signaling, which inhibits JNK-mediated phosphorylation of c-Jun
and protects ovarian cancer cells from apoptosis. Moreover, anal. of The Cancer Genome Atlas data showed that
increased DUSP1 expression was assocd. with decreased overall (P = 0.049) and progression-free (P = 0.0005) survival.
Conclusions: These findings provide a new understanding of the mechanisms by which adrenergic pathways can impair
response to chemotherapy and have implications for cancer management. Clin Cancer Res; 22(7); 1713-24. ©2015
AACR.
~1 Citing
42. Role of extracellular calcitonin gene-related peptide in spinal cord mechanisms of cancer-induced bone pain
By Hansen, Rikke R.; Vacca, Valentina; Pitcher, Thomas; Clark, Anna K.; Malcangio, Marzia
From Pain (2016), 157(3), 666-676. Language: English, Database: CAPLUS, DOI:10.1097/j.pain.0000000000000416
Severe pain is a common and debilitating complication of metastatic bone cancer. Current analgesics provide insufficient
pain relief and often lead to significant adverse effects. In models of cancer-induced bone pain, pathol. sprouting of
sensory fibers at the tumor-bone interface occurs concomitantly with reactive astrocytosis in the dorsal horn of the spinal
cord. We obsd. that calcitonin gene-related peptide (CGRP)-fiber sprouting in the bone was assocd. with an increase in
CGRP content in sensory neuron cell bodies in the dorsal root ganglia (DRG) and increased basal and activity-evoked
release of CGRP from their central terminals in the dorsal horn. Intrathecal administration of a peptide antagonist (α-
CGRP8-37) attenuated referred allodynia in the hind paw ipsilateral to bone cancer. CGRP receptor components (CLR
and RAMP1) were up-regulated in dorsal horn neurons and expressed by reactive astrocytes. In primary cultures of
astrocytes, CGRP incubation led to a concn.-dependent increase of forskolin-induced cAMP prodn., which was
attenuated by pretreatment with CGRP8-37. Furthermore, CGRP induced ATP release in astrocytes, which was
inhibited by CGRP8-37. We suggest that the peripheral increase in CGRP content obsd. in cancer-induced bone pain is
mirrored by a central increase in the extracellular levels of CGRP. This increase in CGRP not only may facilitate
glutamate-driven neuronal nociceptive signaling but also act on astrocytic CGRP receptors and lead to release of ATP.
~2 Citings
43. Post-Translational Regulation of the Glucose-6-Phosphatase Complex by Cyclic Adenosine Monophosphate

Is a Crucial Determinant of Endogenous Glucose Production and Is Controlled by the Glucose-6-Phosphate
Transporter
By Soty, Maud; Chilloux, Julien; Delalande, Francois; Zitoun, Carine; Bertile, Fabrice; Mithieux, Gilles; Gautier-Stein,
Amandine
From Journal of Proteome Research (2016), 15(4), 1342-1349. Language: English, Database: CAPLUS,
DOI:10.1021/acs.jproteome.6b00110
SciFinder® Page 18
The excessive endogenous glucose prodn. (EGP) induced by
glucagon participates in the development of type 2 diabetes.
To further understand this hormonal control, we studied the
short-term regulation by cyclic adenosine monophosphate
(cAMP) of the glucose-6-phosphatase (G6Pase) enzyme,
which catalyzes the last reaction of EGP. In gluconeogenic cell
models, a 1-h treatment by the adenylate cyclase activator
forskolin increased G6Pase activity and glucose prodn.
independently of any change in enzyme protein amt. or G6P
content. Using specific inhibitors or protein overexpression, we
showed that the stimulation of G6Pase activity involved the
protein kinase A (PKA). Results of site-directed mutagenesis,
mass spectrometry analyses, and in vitro phosphorylation
expts. suggested that the PKA stimulation of G6Pase activity
did not depend on a direct phosphorylation of the enzyme.
However, the temp.-dependent induction of both G6Pase
activity and glucose release suggested a membrane-based
mechanism. G6Pase is composed of a G6P transporter
(G6PT) and a catalytic unit (G6PC). Surprisingly, we
demonstrated that the increase in G6PT activity was required
for the stimulation of G6Pase activity by forskolin. Our data
demonstrate the existence of a post-translational mechanism
that regulates G6Pase activity and reveal the key role of G6PT
in the hormonal regulation of G6Pase activity and of EGP.
~0 Citings
44. Dynamic changes in protein interaction between AKAP95 and Cx43 during cell cycle progression of A549
cells
By Chen, Xiaoxuan; Kong, Xiangyu; Zhuang, Wenxin; Teng, Bogang; Yu, Xiuyi; Hua, Suhang; Wang, Su; Liang,
Fengchao; Ma, Dan; Zhang, Suhui; et al
From Scientific Reports (2016), 6, 21224. Language: English, Database: CAPLUS, DOI:10.1038/srep21224
Here we show that A-kinase anchoring protein 95 (AKAP95) and connexin 43 (Cx43) dynamically interact during cell
cycle progression of lung cancer A549 cells. Interaction between AKAP95 and Cx43 at different cell cycle phases was
examd. by tandem mass spectrometry (MS/MS), confocal immunofluorescence microscopy, Western blot, and co-
immunopptn. (Co-IP). Over the course of a complete cell cycle, interaction between AKAP95 and Cx43 occurred in two
stages: binding stage from late G1 to metaphase, and sepg. stage from anaphase to late G1. The binding stage was
further subdivided into complex binding to DNA in interphase and complex sepg. from DNA in metaphase. In late G1,
Cx43 translocated to the nucleus via AKAP95; in anaphase, Cx43 sepd. from AKAP95 and aggregated between two
daughter nuclei. In telophase, Cx43 aggregated at the membrane of the cleavage furrow. After mitosis, Cx43 was
absent from the furrow membrane and was located in the cytoplasm. Binding between AKAP95 and Cx43 was reduced
by N-(2-[P-Bromocinnamylamino]-ethyl)-5-isoquinolinesulfonmide (H89) treatment and enhanced by Forskolin. Dynamic
interaction between AKAP95 and Cx43 varies with cell cycle progression to regulate multiple biol. processes.
~1 Citing
45. Novel effects of FTY720 on perinuclear reorganization of keratin network induced by

sphingosylphosphorylcholine: Involvement of protein phosphatase 2A and G-protein-coupled receptor-12
By Park, Mi Kyung; Park, Soyeun; Kim, Hyun Ji; Kim, Eun Ji; Kim, So Yeon; Kang, Gyeoung Jin; Byun, Hyun Jung;
Kim, Sang Hee; Lee, Ho; Lee, Chang Hoon
From European Journal of Pharmacology (2016), 775, 86-95. Language: English, Database: CAPLUS,
DOI:10.1016/j.ejphar.2016.02.024
SciFinder® Page 19
Sphingosylphosphorylcholine (SPC) evokes perinuclear reorganization of keratin 8 (K8) filaments and regulates the
viscoelasticity of metastatic cancer cells leading to enhanced migration. Few studies have addressed the compds.
modulating the viscoelasticity of metastatic cancer cells. We studied the effects of sphingosine (SPH), sphingosine 1-
phosphate (S1P), FTY720 and FTY720-phosphate (FTY720P) on SPC-induced K8 phosphorylation and reorganization
using Western blot and confocal microscopy, and also evaluated the elasticity of PANC-1 cells by at. force microscopy.
FTY720, FTY720P, SPH, and S1P concn.-dependently inhibited SPC-evoked phosphorylation and reorganization of K8,
and migration of PANC-1 cells. SPC triggered redn. and narrow distribution of elastic const. K and conversely, FTY720
blocked them. A common upstream regulator of JNK and ERK, protein phosphatase 2A (PP2A) expression was reduced
by SPC, but was restored by FTY720 and FTY72P. Butyryl forskolin, a PP2A activator, suppressed SPC-induced K8
phosphorylation and okadaic acid, a PP2A inhibitor, induced K8 phosphorylation. Gene silencing of PP2A also led to K8
phosphorylation, reorganization and migration. We also investigated the involvement of GPR12, a high-affinity SPC
receptor, in SPC-evoked keratin phosphorylation and reorganization. GPR12 siRNA suppressed the SPC-triggered
phosphorylation and reorganization of K8. GPR12 overexpression stimulated keratin phosphorylation and reorganization
even without SPC. FTY720 and FTY720P suppressed the GPR12-induced phosphorylation and reorganization of K8.
The collective data indicates that FTY720 and FTY720P suppress SPC-induced phosphorylation and reorganization of
K8 in PANC-1 cells by restoring the expression of PP2A via GPR12. These findings might be helpful in the development
of compds. that modulate the viscoelasticity of metastatic cancer cells and various SPC actions.
~2 Citings
46. Obesity-induced p53 activation in insulin-dependent and independent tissues is inhibited by beta-adrenergic
agonist in diet-induced obese rats
By Zand, Hamid; Homayounfar, Reza; Cheraghpour, Makan; Jeddi-Tehrani, Mahmood; Ghorbani, Arman; Pourvali,
Katayoun; Soltani, Sama Reza
From Life Sciences (2016), 147, 103-109. Language: English, Database: CAPLUS, DOI:10.1016/j.lfs.2016.01.040
The purpose of this study was to assay the role of beta-adrenergic receptor signaling in the regulation of obesity-induced
p53 in high fat feeding obese rats. The role of beta-adrenergic receptor/cAMP in the regulation of p53 and its
downstream mediators was evaluated by western blot and real-time quant. RT-PCR among diet induced rats. Beta-
adrenergic receptor agonist, isoproterenol, and an adenylate cyclase activator, forskolin, at a single dose significantly
reduced insulin resistance consistent with a decrease in total and phospho-p53 levels in insulin and non-insulin metabolic
target tissues. The decrease of p53 signaling was consistent with the elevation of AKT and subsequent activation.
Obese rats exposed to fasting also exhibited improvement in insulin action despite a slight effect on p53 level. Results of
the present study obviously showed that beta-adrenergic receptor agonist/cAMP prevented obesity-induced p53
activation. Although this effect in metabolic insulin target tissues tempted us to consider them as insulin sensitizers in
obesity-related diabetes, p53 inhibition in non-insulin target tissues warned about the impairment of anti-cancer
mechanisms in obese subjects.
~2 Citings
47. The 4'-hydroxyl group of resveratrol is functionally important for direct activation of PPARα
By Takizawa, Yoshie; Nakata, Rieko; Fukuhara, Kiyoshi; Yamashita, Hiroshi; Kubodera, Hideo; Inoue, Hiroyasu
From PLoS One (2015), 10(3), e0120865/1-e0120865/13. Language: English, Database: CAPLUS,
DOI:10.1371/journal.pone.0120865
Long-term moderate consumption of red wine is assocd. with a reduced risk of developing lifestyle-related diseases such
as cardiovascular disease and cancer. Therefore, resveratrol, a constituent of grapes and various other plants, has
attracted substantial interest. This study focused on one mol. target of resveratrol, the peroxisome proliferator activated
receptor α (PPARα). Our previous study in mice showed that resveratrol-mediated protection of the brain against stroke
requires activation of PPARα; however, the mol. mechanisms involved in this process remain unknown. Here, we
evaluated the chem. basis of the resveratrol-mediated activation of PPARα by performing a docking mode simulation and
examg. the structure-activity relationships of various polyphenols. The results of expts. using the crystal structure of the
PPARα ligand-binding domain and an anal. of the activation of PPARα by a resveratrol analog 4-phenylazophenol (4-
PAP) in vivo indicate that the 4'-hydroxyl group of resveratrol is crit. for the direct activation of PPARα. Activation of
PPARα by 5 µM resveratrol was enhanced by rolipram, an inhibitor of phosphodiesterase (PDE) and forskolin, an
activator of adenylate cyclase. We also found that resveratrol has a higher PDE inhibitory activity (IC50 = 19 µM) than
resveratrol analogs trans-4-hydroxystilbene and 4-PAP (IC50 = 27-28 µM), both of which has only 4'-hydroxyl group,
indicating that this 4'-hydroxyl group of resveratrol is not sufficient for the inhibition of PDE. This result is consistent with
that 10 µM resveratrol has a higher agonistic activity of PPARα than these analogs, suggesting that there is a
feedforward activation loop of PPARα by resveratrol, which may be involved in the long-term effects of resveratrol in vivo.
~0 Citings
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48. Vimentin-mediated steroidogenesis induced by phthalate esters: involvement of DNA demethylation and
nuclear factor κB
By Li, Yuan; Hu, Yanhui; Dong, Congcong; Lu, Hongchao; Zhang, Chang; Hu, Qi; Li, Shifeng; Qin, Heng; Li, Zhong;
Wang, Yubang
Di-Bu phthalate (DBP) and its active metabolite, monobutyl phthalate (MBP) are the most common endocrine disrupting
chems. Many studies indicate that high-doses of DBP and/or MBP exhibit toxicity on testicular function, however, little
attention have been paid to the effects of low levels of DBP/MBP on steroidogenesis. As we all know, the steroidogenic
acute regulatory protein (StAR) is a key regulator involved in the steroidogenesis. Here we found that, in addn. to StAR,
MBP/DBP increased the steroidogenesis by a cytoskeletal protein, vimentin. Briefly, in murine adrenocortical tumor (Y1)
and the mouse Leydig tumor (MLTC-1) cells, vimentin regulated the secretion of progesterone. When these two cells
were exposure to MBP, the DNA demethylation in the vimentin promoter was obsd. In addn., MBP also induced the
activation of nuclear factor kappa B (NF-κB, a transcriptional regulator of vimentin). These two processes improved the
transcriptional elevation of vimentin. Knockdown of NF-κB/vimentin signalling blocked the DBP/MBP-induced
steroidogenesis. These in vitro results were also confirmed via an in vivo model. By identifying a mechanism whereby
DBP/MBP regulates vimentin, our results expand the understanding of the endocrine disrupting potential of phthalate
esters.
~1 Citing
49. Escherichia coli heat-stable enterotoxin mediates Na+/H+ exchanger 4 inhibition involving cAMP in T84
human intestinal epithelial cells
By Beltran, Ana R.; Carraro-Lacroix, Luciene R.; Bezerra, Camila N. A.; Cornejo, Marcelo; Norambuena, Katrina;
Toledo, Fernando; Araos, Joaquin; Pardo, Fabian; Leiva, Andrea; Sanhueza, Carlos; et al
The enterotoxigenic Escherichia coli strains lead to diarrhoea in humans due to heat-labile and heat-stable (STa)
enterotoxins. STa increases Cl-release in intestinal cells, including the human colonic carcinoma T84 cell line, involving
increased cGMP and membrane alkalization due to reduced Na+/H+ exchangers (NHEs) activity. Since NHEs modulate
intracellular pH (pHi), and NHE1, NHE2, and NHE4 are expressed in T84 cells, we characterized the STa a role as
modulator of these exchangers. pHi was assayed by the NH4Cl pulse technique and measured by fluorescence
microscopy in BCECF-preloaded cells. pHi recovery rate (dpHi/dt) was detd. in the absence or presence of 0.25 µmol/L
STa (30 min), 25 µmol/L HOE-694 (concn. inhibiting NHE1 and NHE2), 500 µmol/L sodium nitroprusside (SNP,
spontaneous nitric oxide donor), 100 µmol/L dibutyryl cyclic GMP (db-cGMP), 100 nmol/L H89 (protein kinase A
inhibitor), or 10 µmol/L forskolin (adenylyl cyclase activator). cGMP and cAMP were measured in cell exts. by RIA, and
buffering capacity (βi) and H+ efflux (JH+) was detd. NHE4 protein abundance was detd. by western blotting. STa and
HOE-694 caused comparable redn. in dpHi/dt and JH+ (∼63%), without altering basal pHi (range 7.144-7.172). STa did
not alter βi value in a range of 1.6 pHi units. The dpHi/dt and JH+ was almost abolished (∼94% inhibition) by STa +
HOE-694. STa effect was unaltered by db-cGMP or SNP. However, STa and forskolin increased cAMP level. STa-
decreased dpHi/dt and JH+ was mimicked by forskolin, and STa + HOE-694 effect was abolished by H89. Thus,
incubation of T84 cells with STa results in reduced NHE4 activity leading to a lower capacity of pHi recovery requiring
cAMP, but not cGMP. STa effect results in a causal phenomenon (STa/increased cAMP/increased PKA activity/reduced
NHE4 activity) ending with intracellular acidification that could have consequences in the gastrointestinal cells function
promoting human diarrhoea.
~2 Citings
50. Effects of hydrostatic pressure on carcinogenic properties of epithelia

By Tokuda, Shinsaku; Kim, Young Hak; Matsumoto, Hisako; Muro, Shigeo; Hirai, Toyohiro; Mishima, Michiaki; Furuse,
Mikio
SciFinder® Page 21
The relationship between chronic inflammation and cancer is well known. The inflammation increases the permeability of
blood vessels and consequently elevates pressure in the interstitial tissues. However, there have been only a few
reports on the effects of hydrostatic pressure on cultured cells, and the relationship between elevated hydrostatic
pressure and cell properties related to malignant tumors is less well understood. Therefore, we investigated the effects
of hydrostatic pressure on the cultured epithelial cells seeded on permeable filters. Surprisingly, hydrostatic pressure
from basal to apical side induced epithelial stratification in Madin-Darby canine kidney (MDCK) I and Caco-2 cells, and
cavities with microvilli and tight junctions around their surfaces were formed within the multi-layered epithelia. The
hydrostatic pressure gradient also promoted cell proliferation, suppressed cell apoptosis, and increased transepithelial
ion permeability. The inhibition of protein kinase A (PKA) promoted epithelial stratification by the hydrostatic pressure
whereas the activation of PKA led to suppressed epithelial stratification. These results indicate the role of the hydrostatic
pressure gradient in the regulation of various epithelial cell functions. The findings in this study may provide clues for the
development of a novel strategy for the treatment of the carcinoma.
~0 Citings
51. cAMP signaling increases histone deacetylase 8 expression by inhibiting JNK-dependent degradation via
autophagy and the proteasome system in H1299 lung cancer cells
By Park, Ji-Yeon; Juhnn, Yong-Sung
CAPLUS, DOI:10.1016/j.bbrc.2016.01.049
This study aimed to investigate the roles of autophagy and the ubiquitin-proteasome system in the degrdn. of histone
deacetylase 8 (HDAC8) and to clarify the mechanism by which cAMP signaling regulates this degrdn. CAMP signaling
was activated by treating H1299 non-small cell lung cancer cells with isoproterenol or forskolin/3-isobutyl-1-
methylxanthine, and HDAC8 expression was assessed by western blot anal. The inhibition of autophagy and ubiquitin-
proteasome-dependent degrdn. increased HDAC8 expression. CAMP signaling inhibited JNK activation, which
decreased the phosphorylation of Bcl-2, thereby reducing autophagy, and the phosphorylation of Itch, thereby reducing
ubiquitination. These results suggest that the HDAC8 protein is degraded via autophagy and the ubiquitin-proteasome
system and that cAMP signaling increases HDAC8 protein levels by reducing JNK-mediated autophagy and ubiquitin-
proteasome-dependent degrdn. of the HDAC8 protein in H1299 lung cancer cells.
~1 Citing
52. Investigation of the mechanism and enhancement of the lewis lung carcinoma growth inhibition of
carboxyamidotriazole
By Ju, Rui; Chen, Chen; Chen, Wei; Guo, Lei; Li, Juan; Ye, Caiying; Zhang, Dechang
From Zhonghua Linchuang Yishi Zazhi, Dianziban (2015), 9(10), 107-110. Language: Chinese, Database: CAPLUS,
DOI:10.3877/cma.j.issn.1674-0785.2015.10.028
Objective To investigate the effect of carboxyamidotriazole (CAI) on adenosine 3',5'-cyclic monophosphate specific
phosphodiesterase (cAMP-PDE) in Lewis lung carcinoma (LLC) cells. It was investigated whether adenylate cyclase
(AC) activator Forskolin can enhance the anti-cancer effect of CAI. Methods PDE activity was detd. with the modification
of two-step method of Thompson and Appleman. PDE4 specific inhibitor Rolipram (Rol) was applied as the pos. control.
The live LLC cells were counted with Trypan blue excluding test. Dibutyryl-cAMP was applied to test the sensitivity of
LLC cells to cAMP treatment. Results CAI could inhibit c AMP-PDE activity in LLC cells dose-dependently. The IC50 of
CAI and Rol was 5.62×10-6 mol/L and 1.88×10-8 mol/L, resp. Both Forskolin and db-cAMP could inhibit LLC cell growth
dose-dependently. The inhibition ratio of CAI and Forskolin combination was significantly higher than CAI alone. The
inhibition ratio of 5 µmol/L CAI and 10 µmol/L Forskolin was (32.9±10.3)% and (67.4±3.76)%, resp. Conclusion Inhibition
of cAMP-PDE may be one of the anti-cancer mechanisms of CAI. Its anti-cancer effect can be enhanced by combining
with cAMP elevating drugs such as Forskolin.
~0 Citings
53. Modulation of glucocorticoid, mineralocorticoid and androgen production in H295 cells by Trimesemine, a
mesembrine-rich Sceletium extract
By Swart, A. C.; Smith, C.
From Journal of Ethnopharmacology (2016), 177, 35-45. Language: English, Database: CAPLUS,
DOI:10.1016/j.jep.2015.11.033
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Stress-related illnesses rate among the most prevalent non-fatal diseases globally. With the global trend for consumer
bias towards natural medicine, the Sceletium plant has become more prominent in the field of natural products. Although
potentially useful effects of Sceletium tortuosum on the central nervous system have been reported, limited data is
available on effects of the plant in the peripheral compartment. The current study aimed to elucidate the effect(s) of a
Sceletium ext. (TRI) rich in mesembrine (1% of plant ext. wt./wt.), on adrenal steroid biosynthesis. Steroidogenesis was
assessed basally and in response to stimuli (forskolin, angiotensin II, KCl), in human adrenocortical carcinoma cells
(H295R). Steroid hormone levels were assessed using UPLC-MS/MS. UPLC-MS analyses of TRI identified major
alkaloids ∆7-mesembrenone, mesembrenone and mesembrine. Highest dose TRI treatment (1 mg/mL, 34.5 µM
mesembrine) increased pregnenolone and decreased 16-hydroxyprogesterone levels (both P<0.00001) in forskolin-
stimulated conditions only, suggesting CYP17 enzyme inhibition. This led to significant inhibition of forskolin-assocd.
increases in cortisol levels at the highest dose (P<0.001) and basal cortisol levels across all doses (P<0.0001).
Independently of forskolin, TRI inhibited androstenedione and testosterone prodn. across all doses (both P<0.00001),
suggesting inhibition of 3βHSD and 17βHSD resp. TRI decreased both the angiotensin II- (P<0.05) and forskolin-
induced (P<0.0001) increases in aldosterone prodn. Our data suggest potentially beneficial effects of TRI in the context
of stress and hypertension. These should be further investigated in a whole organism model, while the effects on the
androgenic pathway should also be further elucidated.
~1 Citing
54. Mesenchymal stem cell epidermal differentiation-inducing agent and its application method [Machine
Translation].
By Zhang, Bingqiang; Sun, Hong; Zhang, Gaofeng; Yu, Li
From Faming Zhuanli Shenqing (2015), CN 105112367 A 20151202, Language: Chinese, Database: CAPLUS
[Machine Translation of Descriptors]. The invention belongs to
the tech. field of stem cell induction and differentiation technol.,
in particular to a mesenchymal stem cell differentiation-inducing
agent, and it is made by KGF, BMP-4, PDGF-AB, VEGF, IL-2
and Forskolin. After the addn. of complete medium, the
component concn. is as follows : KGF 20 - 30 µg/L, BMP-4 5 -
10 µg/L, PDGF-AB 5 - 10 µg/L, VEGF 10 - 20 µg/L, IL-2 5 - 10
µg/L, Forskolin 1 - 2mg/L.The mesenchymal stem cell
epidermal differentiation-inducing agent uses cytokines
combined small mol. for efficient induction and differentiation of
mesenchymal stem cells into epidermal cells, greatly improving
the efficiency of induction and differentiation. The cell has
strong activity after induction, without cell transfection, genetic
changes, cancer risk and ethical issues, with high safety.
~0 Citings
55. Methods and compositions for targeting cancer stem cells by activating the Protein Kinase A pathway to
induce a mesenchymal to epithelial transition
By Pattabiraman, Diwakar; Bierie, Brian; Tam, Wai Leong; Weinberg, Robert A.
Aspects of the disclosure relate to methods that involve
activating the Protein Kinase A (PKA) pathway to induce
cancer stem cells (CSCs) to undergo a mesenchymal to
epithelial transition. Methods provided herein are useful, in
some embodiments, because they render CSCs amenable to
treatment with conventional cancer therapies. In some
embodiments, methods are provided that involve assaying PKA
pathway activity to identify compds. that selectively target
CSCs.
~0 Citings
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56. Highly selective one pot synthesis and biological evaluation of novel 3-(allyloxy)-propylidene acetals of
some natural terpenoids
By Devendar, Ponnam; Kumar, Arigari Niranjana; Bethu, M. S.; Zehra, Amtul; Pamanji, R.; Venkateswara Rao, J.;
Tiwari, Ashok Kumar; Sridhar, Balasubramanian; Satya Srinivas, K. V. N.; Kumar, J. Kotesh
From RSC Advances (2015), 5(113), 93122-93130. Language: English, Database: CAPLUS,
DOI:10.1039/C5RA18517C
A series of 3-(allyloxy)-propylidene acetals 1a-6a of some natural terpenoids like andrographolides 1, 2, forskolins 3-5
and arjunolic acid 6 were developed by a novel one pot synthetic strategy using ceric ammonium nitrate as a catalyst.
The method is both chemo and regioselective towards 1,3-acetal formation without affecting other poly functional groups
of terpenoids. O-Allylation is an important functional group transformation for alcs. and the resulting end allylic double
bond may participate in a no. of synthetically useful transformations like olefin metathesis. Acetal of andrographolide 1a
was further converted into 1b, 1c and 1d by dimerization, acetylation and epoxidn. resp. All the synthesized compds.
were screened for in vitro antiproliferative activity against four cancer cell lines B16F10, THP-1, PC-3 and SKOV3.
Derivs. of andrographolide 1a, 1b, 1c and 2a, forskolin 4a and arjunolic acid 6a have shown promising cytotoxicity (IC50 <
10 µg ml-1) in most of the tested cell lines. Also compds. 1b (IC50 0.83 µg ml-1) and 5a (IC50 3.43 µg ml-1) showed
significant α-glucosidase inhibition in an in vitro assay. Structures of all the synthesized compds. were confirmed by
NMR, mass and IR spectral data. A single crystal X-ray anal. of 5a also confirmed the 3-(allyloxy)-propylidene acetal
formation.
~1 Citing
57. A novel 3D high-content assay identifies compounds that prevent fibroblast invasion into tissue surrogates
By Wenzel, Carsten; Otto, Saskia; Prechtl, Stefan; Parczyk, Karsten; Steigemann, Patrick
DOI:10.1016/j.yexcr.2015.10.003
Invasion processes underlie or accompany several pathol. processes but only a limited no. of high-throughput capable
phenotypic models exist to test anti-invasive compds. in vitro. We here evaluated 3D co-cultures as a high-content
phenotypic screening system for fibrotic invasive processes. 3D multicellular spheroids were used as living tissue
surrogates in co-culture with fluorescently labeled lung fibroblasts to monitor invasion processes by automated
microscopy. This setup was used to screen a compd. library contg. 480 known bioactive substances. Identified hits
prevented fibroblast invasion and could be subdivided into two hit classes. First, Prostaglandins were shown to prevent
fibroblast invasion, most likely mediated by the prostaglandin EP2 receptor and generation of cAMP. Addnl., Rho-
assocd. protein kinase (ROCK) inhibitors prevented fibroblast invasion, possibly by inactivation of myosin II. Importantly,
both Prostaglandins and ROCK inhibitors are potential treatment options shown to be effective in in vitro and in vivo
models of fibrotic diseases. This validates the presented novel phenotypic screening approach for the evaluation of
potential inhibitors and the identification of novel compds. with activity in diseases that are assocd. with fibroblast
invasion.
~3 Citings
58. Anticancer agent containing arctigenin and other antitumor component, and side effect-alleviating agent
containing arctigenin
By Esumi, Hiroyasu; Ikeda, Kouji; Tsuchihara, Katsuya; Chiba, Shigeki; Yomoda, Satoshi; Kawashima, Takanori;
Okubo, Toshiki; Tezuka, Yasuhiro; Murata, Kenta
From PCT Int. Appl. (2015), WO 2015156409 A1 20151015, Language: Japanese, Database: CAPLUS
SciFinder® Page 24
The purpose of the present invention is to provide an
anticancer agent for potentiating an antitumor effect, alleviating
side effects, and further extending the survival rate by
concomitant use with a component having an anticancer effect.
An anticancer agent combining arctigenin and a component
other than arctigenin that has an anticancer effect, in which the
anticancer agent may be a combination drug or may be a kit
configured from a formulation contg. arctigenin and a
formulation contg. a component that has an anticancer effect,
and the concomitant use of arctigenin and the component
having an anticancer effect more strongly inhibits tumor growth
and reduces the proportion of cancer stem cells in the tumor,
making it possible to extend the total survival time and to
alleviate side effects caused by the component having an
anticancer effect. Thus, combined administration of
gemcitabine and edible burdock seed ext. contg. arctigenin to
mice bearing Miapaca-2 human pancreatic cancer cells
significantly decreased a rate of cells triple pos. for CD24,
CD44, and ESA (CD326) as markers of cancer stem cells in
the tumor cells.
~1 Citing
59. Activation of the tumor suppressor PP2A emerges as a potential therapeutic strategy for treating prostate
cancer
By Cristobal, Ion; Alonso, Paula Gonzalez; Daoud, Lina; Solano, Esther; Torrejon, Blanca; Manso, Rebeca; Gurpide,
Juan Madoz; Rojo, Federico; Foncillas, Jesus Garcia
From Marine Drugs (2015), 13(6), 3276-3286. Language: English, Database: CAPLUS, DOI:10.3390/md13063276
Protein phosphatase 2A (PP2A) is a tumor suppressor complex that has recently been reported as a novel and highly
relevant mol. target in prostate cancer (PCa). However, its potential therapeutic value remains to be fully clarified. We
treated PC-3 and LNCaP cell lines with the PP2A activators forskolin and FTY720 alone or combined with the PP2A
inhibitor okadaic acid. We examd. PP2A activity, cell growth, prostasphere formation, levels of PP2A phosphorylation,
CIP2A and SET expression, and AKT and ERK activation. Interestingly, both forskolin and FTY720 dephosphorylated
and activated PP2A, impairing proliferation and prostasphere formation and inducing changes in AKT and ERK
phosphorylation. Moreover, FTY720 led to reduced CIP2A levels. Treatment with okadaic acid impaired PP2A activation
thus demonstrating the antitumoral PP2A-dependent mechanism of action of both forskolin and FTY720. Levels of PP2A
phosphorylation together with SET and CIP2A protein expression were studied in 24 PCa patients and both were assocd.
with high Gleason scores and presence of metastatic disease. Altogether, our results suggest that PP2A inhibition could
be involved in PCa progression, and the use of PP2A-activating drugs might represent a novel alternative therapeutic
strategy for treating PCa patients.
~3 Citings
60. Antibodies that bind human cannabinoid 1 (CB1) receptor

By Kretz-Rommel, Anke; Shi, Lei; Ferrini, Roger; Yang, Teddy; Xu, Fei; Campion, Brian
The authors disclose the prepn. and characterization of murine, chimeric, and humanized antibodies that bind human
cannabinoid receptor 1 (CB1). In one example, an anti-CB1 humanized antibody is shown to increase insulin sensitivity
and to reduce triglyceride levels and other cardiovascular risk factors in primates.
~0 Citings
61. MEK Inhibitors Reverse cAMP-Mediated Anxiety in Zebrafish

By Lundegaard, Pia R.; Anastasaki, Corina; Grant, Nicola J.; Sillito, Rowland R.; Zich, Judith; Zeng, Zhiqiang;
Paranthaman, Karthika; Larsen, Anders Peter; Armstrong, J. Douglas; Porteous, David J.; et al
From Chemistry & Biology (Oxford, United Kingdom) (2015), 22(10), 1335-1346. Language: English, Database:
CAPLUS, DOI:10.1016/j.chembiol.2015.08.010
SciFinder® Page 25
Altered phosphodiesterase (PDE)-cAMP (cAMP) activity is frequently assocd. with anxiety disorders, but current
therapies act by reducing neuronal excitability rather than targeting PDE-cAMP-mediated signaling pathways. Here, we
report the novel repositioning of anti-cancer MEK inhibitors as anxiolytics in a zebrafish model of anxiety-like behaviors.
PDE inhibitors or activators of adenylate cyclase cause behaviors consistent with anxiety in larvae and adult zebrafish.
Small-mol. screening identifies MEK inhibitors as potent suppressors of cAMP anxiety behaviors in both larvae and adult
zebrafish, while causing no anxiolytic behavioral effects on their own. The mechanism underlying cAMP-induced anxiety
is via crosstalk to activation of the RAS-MAPK signaling pathway. We propose that targeting crosstalk signaling
pathways can be an effective strategy for mental health disorders, and advance the repositioning of MEK inhibitors as
behavior stabilizers in the context of increased cAMP.
~0 Citings
62. Allocrite Sensing and Binding by the Breast Cancer Resistance Protein (ABCG2) and P-Glycoprotein
(ABCB1)
By Xu, Yanyan; Egido, Estefania; Li-Blatter, Xiaochun; Muller, Rita; Merino, Gracia; Berneche, Simon; Seelig, Anna
From Biochemistry (2015), 54(40), 6195-6206. Language: English, Database: CAPLUS,
DOI:10.1021/acs.biochem.5b00649
The ATP binding cassette (ABC) transporters ABCG2 and
ABCB1 perform ATP hydrolysis-dependent efflux of structurally
highly diverse compds., collectively called allocrites. Whereas
much is known about allocrite-ABCB1 interactions, the chem.
nature and strength of ABCG2-allocrite interactions have not
yet been assessed. We quantified and characterized
interactions of allocrite with ABCG2 and ABCB1 using a set of
39 diverse compds. We also investigated potential allocrite
binding sites based on available transporter structures and
structural models. We demonstrate that ABCG2 binds its
allocrites from the lipid membrane, despite their hydrophilicity.
Hence, binding of allocrite to both transporters is a two-step
process, starting with a lipid-water partitioning step, driven
mainly by hydrophobic interactions, followed by a transporter
binding step in the lipid membrane. We show that binding of
allocrite to both transporters increases with the no. of hydrogen
bond acceptors in allocrites. Scrutinizing the transporter
translocation pathways revealed ample hydrogen bond donors
for allocrite binding. Importantly, the hydrogen bond donor
strength is, on av., higher in ABCG2 than in ABCB1, which
explains the higher measured affinity of allocrite for ABCG2. π-
π Stacking and π-cation interactions play addnl. roles in binding
of allocrite to ABCG2 and ABCB1. With this anal., we
demonstrate that these membrane-mediated weak electrostatic
interactions between transporters and allocrites allow for
transporter promiscuity toward allocrites. The different
sensitivities of the transporters to allocrites' charge and
amphiphilicity provide transporter specificity. In addn., we
show that the different hydrogen bond donor strengths in the
two transporters allow for affinity tuning.
~4 Citings
63. Predicting Activators and Inhibitors of the Breast Cancer Resistance Protein (ABCG2) and P-Glycoprotein
(ABCB1) Based on Mechanistic Considerations
By Egido, Estefania; Muller, Rita; Li-Blatter, Xiaochun; Merino, Gracia; Seelig, Anna
From Molecular Pharmaceutics (2015), 12(11), 4026-4037. Language: English, Database: CAPLUS,
DOI:10.1021/acs.molpharmaceut.5b00463
SciFinder® Page 26
Colocalized in membrane barriers, the ABC transporters
ABCB1 and ABCG2 strongly contribute to multidrug resistance
(MDR). Here we investigate the as yet unknown mechanisms
of activation and inhibition of ABCG2. For this purpose we
measured the ATPase activity of ABCG2 and ABCB1 as a
function of allocrite concn. using a calibration set of 30 diverse
compds. and a validation set of 23 compds. We demonstrate
that ABCG2 is activated at low and inhibited at high allocrite
concns., yielding bell-shaped activity curves. With an ATP
regeneration assay we prove that the inhibitory part is indeed
due to a decrease in activity because of high allocrite load in
the transporter. However, inhibition is only obsd. if the
membrane soly. of allocrites is sufficiently high. The concns. of
half-max. activation and inhibition are at least 10-fold lower for
ABCG2 than for ABCB1. Because ABCG2 binds its allocrites
with higher affinity than ABCB1, it can ext. hydrophilic,
nonamphiphilic, and highly charged compds. out of the lipid
membrane, typically exhibiting low lipid-water partition coeffs.,
but is inhibited by hydrophobic, amphiphilic, and moderately
charged compds., with high lipid-water partition coeffs. In
contrast, ABCB1 is barely interacting with hydrophilic compds.,
but is activated by hydrophobic compds. We show that
hydrophobicity, amphiphilicity, and charge have a dual role;
they predict, on the one hand, allocrites' lipid-water partition
coeff. and, on the other hand, the transporters' preference for
the chem. nature of allocrites. Parameters reflecting
hydrophobicity, amphiphilicity, and charge are therefore
sufficient for differentiating between allocrites, activators, and
inhibitors of ABCB1 and ABCG2.
~8 Citings
64. Low-Dose Endothelial Monocyte-Activating Polypeptide-II Induces Blood-Tumor Barrier Opening Via the
cAMP/PKA/Rac1 Pathway
By Li, Zhen; Liu, Xiao-bai; Liu, Yun-hui; Xue, Yi-xue; Liu, Jing; Teng, Hao; Xi, Zhuo; Yao, Yi-long
From Journal of Molecular Neuroscience (2016), 58(2), 153-161. Language: English, Database: CAPLUS,
DOI:10.1007/s12031-015-0649-8
Previous studies have demonstrated that low-dose endothelial monocyte-activating polypeptide-II (EMAP-II) induces
blood-tumor barrier (BTB) hyperpermeability via both paracellular and transcellular pathways. In a recent study, we
revealed that cyclic adenosine monophosphate (cAMP)/protein kinase A (PKA)-dependent signaling pathway is involved
in EMAP-II-induced BTB hyperpermeability. This study further investigated the exact mechanisms through which the
cAMP/PKA-dependent signaling pathway affects EMAP-II-induced BTB hyperpermeability. In an in vitro BTB model, low-
dose EMAP-II (0.05 nM) induced a significant decrease in Rac1 activity in rat brain microvascular endothelial cells
(RBMECs). Pretreatment with forskolin to elevate intracellular cAMP concn. completely blocked EMAP-II-induced
inactivation of Rac1. Besides, pretreatment with 6Bnz-cAMP to activate PKA partially attenuated EMAP-II-induced Rac1
inactivation. Moreover, 6Bnz-cAMP pretreatment significantly diminished EMAP-II-induced changes in BTB permeability,
myosin light chain (MLC) phosphorylation, expression and distribution of ZO-1, and actin cytoskeleton arrangement in
RBMECs. These effects of 6Bnz-cAMP were completely blocked in the presence of NSC-23766 (the specific inhibitor of
Rac1). In conclusion, this study demonstrates that low-dose EMAP-II induces BTB hyperpermeability via the
cAMP/PKA/Rac1 signaling pathway.
~0 Citings
65. Methods and compositions for diagnosing and treating loss and/or distortion of taste or smell
By Henkin, Robert I.
Disclosed herein are methods for diagnosing a subject with loss and/or distortion of taste or smell, e.g., hyposmia,
dysosmia, anosmia, phantosmia, hypogeusia, dysgeusia, phantogeusia, and/or ageusia. Also disclosed herein are
methods and compns. for treating a subject for loss and/or distortion of taste or smell, e.g., hyposmia, dysosmia,
anosmia, phantosmia, hypogeusia, dysgeusia, phantogeusia, and/or ageusia.
SciFinder® Page 27
~0 Citings
66. Selective Generation of Dopaminergic Precursors from Mouse Fibroblasts by Direct Lineage Conversion
By Tian, Changhai; Li, Yuju; Huang, Yunlong; Wang, Yongxiang; Chen, Dapeng; Liu, Jinxu; Deng, Xiaobei; Sun, Lijun;
Anderson, Kristi; Qi, Xinrui; et al
Degeneration of midbrain dopaminergic (DA) neurons is a key pathol. event of Parkinson's disease (PD). Limited adult
dopaminergic neurogenesis has led to novel therapeutic strategies such as transplantation of dopaminergic precursors
(DPs). However, this strategy is currently restrained by a lack of cell source, the tendency for the DPs to become a glial-
restricted state, and the tumor formation after transplantation. Here, we demonstrate the direct conversion of mouse
fibroblasts into induced DPs (iDPs) by ectopic expression of Brn2, Sox2 and Foxa2. Besides expression with neural
progenitor markers and midbrain genes including Corin, Otx2 and Lmx1a, the iDPs were restricted to dopaminergic
neuronal lineage upon differentiation. After transplantation into MPTP-lesioned mice, iDPs differentiated into DA
neurons, functionally alleviated the motor deficits, and reduced the loss of striatal DA neuronal axonal termini.
Importantly, no iDPs-derived astroctyes and neoplasia were detected in mouse brains after transplantation. We propose
that the iDPs from direct reprogramming provides a safe and efficient cell source for PD treatment.
~6 Citings
67. Method for providing information of damaged DNA repairing speed

By Kang, Tae Hong; Park, Jeong Min; Choi, Ji Ye; Lim, Seon Hui
From Repub. Korea (2015), KR 1542324 B1 20150805, Language: Korean, Database: CAPLUS
The invention relates to method for information providing
information of damaged DNA repairing speed which is
ATR(Ataxia telangiectasia and Rad3 related) activating time
according to cryptochrome expression. It is about the
information of the repairing speed of damaged DNA, which can
be utilized to det. the time when the anti-cancer side effect is
less and decide the anti-cancer drug dose timing.
~0 Citings
68. G-protein coupled receptors in hedgehog signaling and identifying TPRA40 antagonists and agonists for
therapeutic use
By Singh, Jaskirat; Scales, Susanna J.
The disclosure provides methods for identifying TPRA40 antagonists and agonists. The method further provides
methods for inhibiting hedgehog signaling and/or inhibiting unwanted cell proliferation, such as unwanted cell proliferation
caused, in whole or in part, by hyperactive hedgehog signaling, using a TPRA40 antagonist. In certain embodiments of
the disclosure, the TPRA40 antagonist is a polynucleotide mol. or antibody that inhibits the expression of TPRA40.
~0 Citings
69. Posttranscriptional regulation of T-type Ca2+ channel expression by interleukin-6 in prostate cancer cells
SciFinder® Page 28
By Weaver, Erika M.; Zamora, Francis J.; Hearne, Jennifer L.; Martin-Caraballo, Miguel
From Cytokine+ (2015), 76(2), 309-320. Language: English, Database: CAPLUS, DOI:10.1016/j.cyto.2015.07.004
At early stages, the growth of prostate cancers is androgen dependent. At later stages, however, the growth of prostate
cancers becomes androgen independent, which leads to an increase in mortality. The switch to an androgen-refractory
state is assocd. with neuroendocrine differentiation (NED) of prostate cancer cells. Several factors including interleukin-6
(IL-6) and increased cAMP prodn. promote NED of prostate cancer cells. In this work we investigated whether IL-6
evoked NED of LNCaP cells results in a significant change in T-type Ca2+ channel expression in comparison to non-
stimulated LNCaP cells. T-type Ca2+ channel subunit Cav3.2 expression was studied using PCR anal., western blot and
whole cell recordings. Tubulin IIIβ expression and neurite-like morphol. was assessed to investigate the role of T-type
Ca2+ channels in the differentiation of prostate cancer cells. Treatment of LNCaP cells with IL-6 for 4 days evokes
considerable morphol. and biochem. changes consistent with NED. Transcripts of the T-type Ca2+ channel subunit
Cav3.2 but not Cav3.1 or Cav3.3 are detected in IL-6 stimulated cells. Real time PCR anal. of IL-6 stimulated cells
indicates no significant change in Cav3.2 mRNA expression in comparison to non-stimulated cells. LNCaP cells
stimulated with IL-6 show a threefold increase in T-type Ca2+ channel subunit Cav3.2 protein expression, suggesting that
channel expression is upregulated by a posttranscriptional mechanism. Electrophysiol. recordings reveal that increased
Cav3.2 protein expression following IL-6 stimulation of LNCaP cells does not result in increased expression of functional
channels in the membrane. Functional expression of Cav3.2 channels in LNCaP cells is facilitated by co-stimulation with
IL-6 and the cAMP-stimulating agent, forskolin (FSK). Inhibition of T-type Ca2+ channel activity in IL-6 stimulated LNCaP
cells prevents the development of morphol. characteristics consistent with NED. These results indicate that the
functional expression of T-type Ca2+ channels is regulated by the interplay of multiple factors in LNCaP cells.
~3 Citings
70. Inhibition of breast cancer cell migration by activation of cAMP signaling

By Dong, Hongli; Claffey, Kevin P.; Brocke, Stefan; Epstein, Paul M.
From Breast Cancer Research and Treatment (2015), 152(1), 17-28. Language: English, Database: CAPLUS,
DOI:10.1007/s10549-015-3445-9
Almost all deaths from breast cancer arise from metastasis of the transformed cells to other sites in the body. Hence,
uncovering a means of inhibiting breast cancer cell migration would provide a significant advance in the treatment of this
disease. Stimulation of the cAMP signaling pathway has been shown to inhibit migration and motility of a no. of cell
types. A very effective way of selectively stimulating cAMP signaling is through inhibition of cyclic nucleotide
phosphodiesterases (PDEs). Therefore, we examd. full expression profiles of all known PDE genes at the mRNA and
protein levels in four human breast cancer cell lines and eight patients' breast cancer tissues. By these analyses,
expression of almost all PDE genes was seen in both cell lines and tissues. In the cell lines, appreciable expression was
seen for PDEs 1C, 2A, 3B, 4A, 4B, 4D, 5A, 6B, 6C, 7A, 7B, 8A, 9A, 10A, and 11A. In patients' tissues, appreciable
expression was seen for PDEs 1A, 3B, 4A, 4B, 4C, 4D, 5A, 6B, 6C, 7A, 7B, 8A, 8B, and 9A. PDE8A mRNA in particular
is prominently expressed in all cell lines and patients' tissue samples examd. We show here that stimulation of cAMP
signaling with cAMP analogs, forskolin, and PDE inhibitors, including selective inhibitors of PDE3, PDE4, PDE7, and
PDE8, inhibit aggressive triple neg. MDA-MB-231 breast cancer cell migration. Under the same conditions, these agents
had little effect on breast cancer cell proliferation. This study demonstrates that PDE inhibitors inhibit breast cancer cell
migration, and thus may be valuable therapeutic targets for inhibition of breast cancer metastasis. Since PDE8A is
expressed in all breast cancer samples, and since dipyridamole, which inhibits PDE8, and PF-04957325, a selective
PDE8 inhibitor, both inhibit migration, it suggests that PDE8A may be a valuable novel target for treatment of this
disease.
~4 Citings
71. Method and medium for neuronal stem cell differentiation

By Kemp, Paul; Allen, Nicholas
SciFinder® Page 29
The invention relates to a neural cell differentiation medium for
the prodn. of neural cells from at least one precursor cell
capable of lineage specific differentiation, a method for neural
cell differentiation using said medium and the use of said
medium and said method for lineage specific differentiation of
at least one precursor cell towards a neural cell fate.
~0 Citings
72. Advances in the analytical methodologies: Profiling steroids in familiar pathways-challenging dogmas
By Bloem, Liezl M.; Storbeck, Karl-Heinz; Swart, Pieter; du Toit, Therina; Schloms, Lindie; Swart, Amanda C.
From Journal of Steroid Biochemistry and Molecular Biology (2015), 153, 80-92. Language: English, Database:
CAPLUS, DOI:10.1016/j.jsbmb.2015.04.009
The comprehensive evaluation of the adrenal steroidogenic pathway, given its complexity, requires methodol. beyond the
std. techniques currently employed. Advances in LC-MS/MS, coupled with in vitro cell models that produce all the steroid
metabolites of the mineralo-, glucocorticoid and androgen arms, present a powerful approach for the comprehensive
evaluation of adrenal steroidogenesis in response to compds. of interest including bioactives, drug treatments and
endocrine disrupting compds. UHPLC-MS/MS anal. of steroid panels in forskolin, angiotensin II and K+ stimulated
H295R cells provides a snapshot of their effect on intermediates and end products of adrenal steroidogenesis. The
impact of full steroid panel evaluations by LC- and GC-MS/MS extends to clin. profiling with the characterization of
normal pediatric steroid ref. ranges in sexual development and of disease-specific profiles improving diagnosis and sub
classification. Comprehensive analyses of steroid profiles may potentially improve patient outcomes together with the
application of treatments specifically suited to clin. subgroups. LC-MS/MS and GC-MS/MS applications in the analyses
of comprehensive steroid panels are demonstrated in clin. conditions such as congenital adrenal hyperplasia in
newborns requiring accurate diagnoses and in predicting metabolic risk in polycystic ovary syndrome patients. Most
notable perhaps is the impact of LC-MS/MS evaluations on our understanding of the basic biochem. of steroidogenesis
with the detection of the long forgotten adrenal steroid, 11β-hydroxyandrostenedione, at significant levels. The
characterization of its metab. to androgen receptor ligands in the LNCaP prostate cancel cell model, specifically within
the context of recurring prostate cancer, lends new perspectives to old dogmas. We demonstrate that UHPLC-MS/MS
has enabled the analyses of novel metabolites of the enzymes, SRD5A, 11βHSD and 17βHSD, in LNCaP cells.
Undoubtedly, the continuous advances in the anal. methodologies used for steroid profiling and quantification will give
impetus to the unraveling of the remaining enigmas, old and new, of both hormone biosynthesis and metab.
~9 Citings
73. Perfluorooctyl Iodide Stimulates Steroidogenesis in H295R Cells via a Cyclic Adenosine Monophosphate
Signaling Pathway
By Wang, Chang; Ruan, Ting; Liu, Jiyan; He, Bin; Zhou, Qunfang; Jiang, Guibin
From Chemical Research in Toxicology (2015), 28(5), 848-854. Language: English, Database: CAPLUS,
DOI:10.1021/tx5004563
SciFinder® Page 30
Perfluorinated iodine alkanes (PFIs) are used widely in the org.
fluorine industry. Increased prodn. of PFIs has caused
environmental health concerns. To evaluate the potential
endocrine-disrupting effect of PFIs, the authors investigated the
effects of perfluorooctyl iodide (PFOI) on steroidogenesis in
human adrenocortical carcinoma cells (H295R). Levels of
aldosterone, cortisol, 17β-estradiol, and testosterone were
measured in H295R culture medium upon treatment with
perfluorooctanoic acid (PFOA) and PFIs. Expression of 10
steroidogenic genes (StAR, HMGR, CYP11A1, 3βHSD2,
17βHSD, CYP17, CYP21, CYP11B1, CYP11B2, and CYP19)
was measured by real-time polymerase chain reaction. Levels
of cAMP and adenylate cyclase (AC) activity were measured to
understand the underlying mechanism of steroidogenic
perturbations. Levels of prodn. of aldosterone, cortisol, and
17β-estradiol were elevated significantly, and the level of
testosterone generation decreased upon treatment with 100
µM PFOI. Similar to the effect induced by forskolin (AC
activator), expression of all 10 genes involved in the synthesis
of steroid hormones was upregulated significantly upon
exposure to 100 µM PFOI. PFOA had no effect on steroid
hormone prodn. or steroidogenic gene expression even though
it is highly structurally similar with PFOI. Therefore, the
terminal -CF2I group in PFOI could be a crit. factor for
mediation of steroidogenesis. PFOI increased AC activity and
cAMP levels in H295R cells, which implied an underlying
mechanism for the disturbance of steroidogenesis. These data
suggest that PFOI may act as an AC activator, thereby
stimulating steroidogenesis by activating a cAMP signaling
pathway.
~7 Citings
74. Salt-Inducible Kinase 2 Regulates Mitotic Progression and Transcription in Prostate Cancer
By Bon, Helene; Wadhwa, Karan; Schreiner, Alexander; Osborne, Michelle; Carroll, Thomas; Ramos-Montoya,
Antonio; Ross-Adams, Helen; Visser, Matthieu; Hoffmann, Ralf; Ahmed, Ahmed Ashour; et al
From Molecular Cancer Research (2015), 13(4), 620-635. Language: English, Database: CAPLUS, DOI:10.1158/1541-
7786.MCR-13-0182-T
Salt-inducible kinase 2 (SIK2) is a multifunctional kinase of the AMPK family that plays a role in CREB1-mediated gene
transcription and was recently reported to have therapeutic potential in ovarian cancer. The expression of this kinase
was investigated in prostate cancer clin. specimens. Interestingly, auto-antibodies against SIK2 were increased in the
plasma of patients with aggressive disease. Examn. of SIK2 in prostate cancer cells found that it functions both as a
pos. regulator of cell-cycle progression and a neg. regulator of CREB1 activity. Knockdown of SIK2 inhibited cell growth,
delayed cell-cycle progression, induced cell death, and enhanced CREB1 activity. Expression of a kinase-dead mutant
of SIK2 also inhibited cell growth, induced cell death, and enhanced CREB1 activity. Treatment with a small-mol. SIK2
inhibitor (ARN-3236), currently in preclin. development, also led to enhanced CREB1 activity in a dose- and time-
dependent manner. Because CREB1 is a transcription factor and proto-oncogene, it was posited that the effects of SIK2
on cell proliferation and viability might be mediated by changes in gene expression. To test this, gene expression array
profiling was performed and while SIK2 knockdown or overexpression of the kinase-dead mutant affected established
CREB1 target genes; the overlap with transcripts regulated by forskolin (FSK), the adenylate cyclase/CREB1 pathway
activator, was incomplete. Implications: This study demonstrates that targeting SIK2 genetically or therapeutically will
have pleiotropic effects on cell-cycle progression and transcription factor activation, which should be accounted for when
characterizing SIK2 inhibitors. Mol Cancer Res; 13(4); 620-35. ©2014 AACR.
~6 Citings
75. Cyclic AMP signaling reduces sirtuin 6 expression in non-small-cell lung cancer cells by promoting
ubiquitin-proteasomal degradation via inhibition of the Raf-MEK-ERK (Raf/Mitogen-activated Extracellular
Signal-regulated Kinase/Extracellular Signal-regulated Kinase) Pathway
By Kim, Eui-Jun; Juhnn, Yong-Sung
From Journal of Biological Chemistry (2015), 290(15), 9604-9613. Language: English, Database: CAPLUS,
DOI:10.1074/jbc.M114.633198
SciFinder® Page 31
The cAMP signaling system regulates various cellular functions, including metab., gene expression, and death. Sirtuin 6
(SIRT6) removes acetyl groups from histones and regulates genomic stability and cell viability. We hypothesized that
cAMP modulates SIRT6 activity to regulate apoptosis. Therefore, we examd. the effects of cAMP signaling on SIRT6
expression and radiation-induced apoptosis in lung cancer cells. cAMP signaling in H1299 and A549 human non-small
cell lung cancer cells was activated via the expression of constitutively active Gαs plus treatment with prostaglandin E2
(PGE2), isoproterenol, or forskolin. The expression of sirtuins and signaling mols. were analyzed by Western blotting.
Activation of cAMP signaling reduced SIRT6 protein expression in lung cancer cells. cAMP signaling increased the
ubiquitination of SIRT6 protein and promoted its degrdn. Treatment with MG132 and inhibiting PKA with H89 or with a
dominant-neg. PKA abolished the cAMP-mediated redn. in SIRT6 levels. Treatment with PGE2 inhibited c-Raf activation
by increasing inhibitory phosphorylation at Ser-259 in a PKA-dependent manner, thereby inhibiting downstream MEK-
ERK signaling. Inhibiting ERK with inhibitors or with dominant-neg. ERKs reduced SIRT6 expression, whereas activation
of ERK by constitutively active MEK abolished the SIRT6-depleting effects of PGE2. cAMP signaling also augmented
radiation-induced apoptosis in lung cancer cells. This effect was abolished by exogenous expression of SIRT6. It is
concluded that cAMP signaling reduces SIRT6 expression by promoting its ubiquitin-proteasome-dependent degrdn., a
process mediated by the PKA-dependent inhibition of the Raf-MEK-ERK pathway. Reduced SIRT6 expression mediates
the augmentation of radiation-induced apoptosis by cAMP signaling in lung cancer cells.
~7 Citings
76. DuOx2 promoter regulation by hormones, transcriptional factors and the coactivator TAZ
By Cardoso-Weide, L. C.; Cardoso-Penha, R. C.; Costa, M. W.; Ferreira, A. C. F.; Carvalho, D. P.; Santisteban, P. S.
From European Thyroid Journal (2015), 4(1), 6-13. Language: English, Database: CAPLUS, DOI:10.1159/000379749
The prodn. of H2O2, which is essential to thyroid hormone synthesis, involves two NADPH oxidases: dual oxidase 1 and
2 (DuOx1 and DuOx2). A functional study with human DuOx genes and their 5'-flanking regions showed that DuOx1 and
-2 promoters are different from thyroid-specific gene promoters. Furthermore, their transcriptional activities are not
restricted to thyroid cells. While regulation of Tg (thyroglobulin) and TPO (thyroperoxidase) expression have been
extensively studied, DuOx2 promoter regulation by hormones and transcriptional factors need to be more explored.
Herein we investigated the role of TSH, insulin and insulin-like growth factor 1 (IGF-1), as well as the cAMP effect on
DuOx2 promoter (ptx41) activity in transfected rat thyroid cell lines (PCCL3). We also assessed DuOx2 promoter activity
in the presence of transcriptional factors crucial to thyroid development such as TTF-1 (thyroid transcription factor 1),
PAX8, CREB, DREAM, Nkx2.5 and the coactivator TAZ in HeLa and HEK 293T-transfected cells. Our results show that
TSH and forskolin, which increase cAMP in thyroid cells, stimulated DuOx2 promoter activity. IGF-1 led to pronounced
stimulation, while insulin induction was not statistically different from DuOx2 promoter basal activity. All transcriptional
factors selected for this work and coactivator TAZ, except DREAM, stimulated DuOx2 promoter activity. Moreover,
Nkx2.5 and TAZ synergistically increased DuOx2 promoter activity. In conclusion, we show that DuOx2 expression is
regulated by hormones and transcription factors involved in thyroid organogenesis and carcinogenesis, reinforcing the
importance of the control of H2O2 generation in the thyroid.
~1 Citing
77. Growth arrest and forced differentiation of human primary glioblastoma multiforme by a novel small
molecule
By Kang, Tae-Wook; Choi, Soon Won; Yang, Se-Ran; Shin, Tae-Hoon; Kim, Hyung-Sik; Yu, Kyung-Rok; Hong, In-Sun;
Ro, Seonggu; Cho, Joong Myung; Kang, Kyung-Sun
Glioblastoma multiforme is the most common malignant brain tumor in adults, with an av. survival of less than one year
due to its resistance to therapy. Recent studies reported that GBM initiates from CD133-expressing cancer stem cells
(CSC). However, the efficacy of CSC targeting is limited. A newly developed approach in cancer treatment is the forced
differentiation of cancer cells. Here, we show that the treatment of the novel small mol., CG500354, into CD133-
expressing human primary GBM cells induces growth arrest by cell cycle regulators, p53, p21, p27 and phase-specific
cyclins, and neural differentiation, as confirmed by neural progenitor/precursor markers, nestin, GFAP and Tuj1. When
GBM-derived cells caused the tumors in NOD/SCID mice, CG500354 induced GBM-derived cells differentiation into Tuj1
and GFAP expressing cells. We next demonstrated that CG500354 plays a tumor-suppressive role via cAMP/CREB
signaling pathway. CG500354 increases not only the extracellular cAMP level but also the protein level of PKA and
CREB. Addnl., both mimetic substances, Forskolin and Rolipram, revealed comparable results with CG500354. Our
findings indicate that induction of growth arrest and neural differentiation via cAMP/CREB signaling pathway by
CG500354 treatment suggests the novel targeting of PDE4D in the development of new drugs for brain tumor therapy.
~6 Citings

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78. Exogenous norepinephrine attenuates the efficacy of sunitinib in a mouse cancer model
By Deng, Guo-Hua; Liu, Jie; Zhang, Jie; Wang, Ying; Peng, Xing-Chen; Wei, Yu-Quan; Jiang, Yu
From Journal of Experimental & Clinical Cancer Research (2014), 33, 21/1-21/12, 12. Language: English, Database:
CAPLUS, DOI:10.1186/1756-9966-33-21
Background: Sunitinib alone exhibits satisfactory efficacy in several mouse homografts and xenografts but unsatisfactory
efficacy in many kinds of solid tumors in clinic. Different from animals, receiving a diagnosis of cancer impacts chronic
stress on patients. Here, we examine whether norepinephrine (NE), one of the most potent stress related hormones,
leads to the difference in the efficacy of sunitinib between clin. and preclin. trials. Methods: The influence of NE on
mouse melanoma B16F1 cells under sunitinib was evaluated in vitro and in vivo. The Β-AR/cAMP/PKA (Β-
adrenoceptor/cyclic adenosine monophosphate/protein kinase A) signaling pathway was also evaluated in human lung
adenocarcinoma cells. Results: We found that NE upregulated the expression of VEGF, IL-8 and IL-6 in vitro and
stimulated tumor growth in vivo, which was mediated by Β-AR/cAMP/PKA signaling pathway and could be inhibited by
propranolol, a Β-blocker for hypertension for decades. Conclusions: This research indicates exogenous norepinephrine
attenuates the efficacy of sunitinib, and a combination of sunitinib and propranolol might be suggested as a new strategy
in solid tumor in clinic.
~2 Citings
79. cAMP signaling inhibits radiation-induced ATM phosphorylation leading to the augmentation of apoptosis in
human lung cancer cells
By Cho, Eun-Ah; Kim, Eui-Jun; Kwak, Sahng-June; Juhnn, Yong-Sung
From Molecular Cancer (2014), 13, 36/1-36/15, 15. Language: English, Database: CAPLUS, DOI:10.1186/1476-4598-
13-36
Background: The ataxia-telangiectasia mutated (ATM) protein kinase plays a central role in coordinating the cellular
response to radiation-induced DNA damage. cAMP signaling regulates various cellular responses including metab. and
gene expression. This study aimed to investigate the mechanism through which cAMP signaling regulates ATM
activation and cellular responses to ionizing radiation in lung cancer cells. Methods: Lung cancer cells were transfected
with constitutively active stimulatory G protein (GαsQL), and irradiated with γ-rays. The phosphorylation of ATM and
protein phosphatase 2A was analyzed by western blotting, and apoptosis was assessed by western blotting, flow
cytometry, and TUNNEL staining. The promoter activity of NF-κB was detd. by dual luciferase reporter assay. BALB/c
mice were treated with forskolin to assess the effect in the lung tissue. Results: Transient expression of GαsQL
significantly inhibited radiation-induced ATM phosphorylation in H1299 human lung cancer cells. Treatment with okadaic
acid or knock down of PP2A B56δ subunit abolished the inhibitory effect of Gαs on radiation-induced ATM
phosphorylation. Expression of GαsQL increased phosphorylation of the B56δ and PP2A activity, and inhibition of PKA
blocked Gαs-induced PP2A activation. GαsQL enhanced radiation-induced cleavage of caspase-3 and PARP and
increased the no. of early apoptotic cells. The radiation-induced apoptosis was increased by inhibition of NF-κB using
PDTC or inhibition of ATM using KU55933 or siRNA against ATM. Pretreatment of BALB/c mice with forskolin stimulated
phosphorylation of PP2A B56δ, inhibited the activation of ATM and NF-κB, and augmented radiation-induced apoptosis
in the lung tissue. GαsQL expression decreased the nuclear levels of the p50 and p65 subunits and NF-κB-dependent
activity after γ-ray irradn. in H1299 cells. Pretreatment with prostaglandin E2 or isoproterenol increased B56δ
phosphorylation, decreased radiation-induced ATM phosphorylation and increased apoptosis. Conclusions: cAMP
signaling inhibits radiation-induced ATM activation by PKA-dependent activation of PP2A, and this signaling mechanism
augments radiation-induced apoptosis by reducing ATM-dependent activation of NF-κB in lung cancer cells.
~3 Citings
80. An integrated microfluidic probe for multiplexed single cell kinase kinase activity measurement
By Sarkar, Aniruddh; Kolitz, Sarah; Lauffenburger, Douglas A.; Han, Jongyoon
Edited By:Fujii, Teruo
From International Conference on Miniaturized Systems for Chemistry and Life Sciences, 16th, Okinawa, Japan, Oct.
28-Nov. 1, 2012 (2014), 1, 416-418. Language: English, Database: CAPLUS
We present an integrated microfluidic probe that enables the measurement of multiple kinase activities in selected
phenotypically distinct single cells from large cell populations on std. tissue culture platforms. The contents of a cell are
captured without disturbing its extracellular context by creating a small lysis zone at the probe tip by hydrodynamic
confinement. Pneumatic micro-valves are then used to sep. and mix the captured lysate into different assay mixts. in
sep. small vol. chambers for a fluorimetric assay. We demonstrate here the ability to simultaneously measure the activity
of three kinases: Akt, MAPKAPK2, PKA and a loading control enzyme, GAPDH, from a single cell. This single cell assay
platform enables the correlation of cellular phenotype to intracellular biochem. state at the single cell level and hence can
provide a clearer understanding of cell behavior in heterogeneous cell populations.
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~0 Citings
81. Inhibitory effects of prostaglandin E2 on collagen synthesis and cell proliferation in human stellate cells
from pancreatic head adenocarcinoma
By Pomianowska, Ewa; Sandnes, Dagny; Grzyb, Krzysztof; Schjoelberg, Aasa R.; Aasrum, Monica; Tveteraas, Ingun
H.; Tjomsland, Vegard; Christoffersen, Thoralf; Gladhaug, Ivar P.
From BMC Cancer (2014), 14, 413/1-413/27, 27 pp.. Language: English, Database: CAPLUS, DOI:10.1186/1471-
2407-14-413
Background: Several studies have described an increased cyclooxygenase-2 (COX-2) expression in pancreatic cancer,
but the role of COX-2 in tumor development and progression is not clear. The aim of the present study was to examine
expression of COX-2 in cancer cells and stromal cells in pancreatic cancer specimens, and to explore the role of PGE2 in
pancreatic stellate cell proliferation and collagen synthesis. Methods: Immunohistochem. and immunofluorescence was
performed on slides from whole sections of tissue blocks using antibodies against COX-2 and α-smooth muscle actin
(αSMA). Pancreatic stellate cells (PSC) were isolated from surgically resected tumor tissue by the outgrowth method.
Cells were used between passages 4 and 8. Collagen synthesis was detd. by [3H]-proline incorporation, or by enzyme
immunoassay measurement of collagen C-peptide. DNA synthesis was measured by incorporation of [3H]-thymidine in
DNA. CAMP (cAMP) was detd. by RIA. Collagen 1A1 mRNA was detd. by RT-qPCR. Results: Immunohistochem.
staining showed COX-2 in pancreatic carcinoma cells, but not in stromal cells. All tumors showed pos. staining for αSMA
in the fibrotic stroma. Cultured PSC expressed COX-2, which could be further induced by interleukin-1β (IL-1β),
epidermal growth factor (EGF), thrombin, and PGE2, but not by transforming growth factor-β1 (TGFβ). Indirect coculture
with the adenocarcinoma cell line BxPC-3, but not HPAFII or Panc-1, induced COX-2 expression in PSC. Treatment of
PSC with PGE2 strongly stimulated cAMP accumulation, mediated by EP2 receptors, and also stimulated
phosphorylation of extracellular signal-regulated kinase (ERK). Treatment of PSC with PGE2 or forskolin suppressed
both TGFβ-stimulated collagen synthesis and PDGF-stimulated DNA synthesis. Conclusions: The present results show
that COX-2 is mainly produced in carcinoma cells and suggest that the cancer cells are the main source of PGE22 in
pancreatic tumors. PGE2 exerts a suppressive effect on proliferation and fibrogenesis in pancreatic stellate cells. These
effects of PGE2 are mediated by the cAMP pathway and suggest a role of EP2 receptors.
~2 Citings
82. Prostaglandin E2 promotes the cell growth and invasive ability of hepatocellular carcinoma cells by
upregulating c-Myc expression via EP4 receptor and the PKA signaling pathway
By Xia, Shukai; Ma, Juan; Bai, Xiaoming; Zhang, Hai; Cheng, Shanyu; Zhang, Min; Zhang, Li; Du, Mingzhan; Wang,
Yipin; Li, Hai; et al
From Oncology Reports (2014), 32(4), 1521-1530. Language: English, Database: CAPLUS, DOI:10.3892/or.2014.3393
Hepatocellular carcinoma (HCC) represents a major health problem worldwide. Prostaglandin E2 (PGE2), the
predominant product of cyclooxygenase-2, has been implicated in hepatocarcinogenesis. However, the underlying mol.
mechanisms remain to be further elucidated. C-myc, a cellular proto-oncogene, is activated or overexpressed in many
types of human cancer, including HCC. The present study was designed to investigate the internal relationship and mol.
mechanisms between PGE2 and c-Myc in HCC, and to define its role in HCC cell growth and invasion. Our results
showed that PGE2 significantly upregulated c-Myc expression at both the mRNA and protein levels, and knockdown of c-
Myc blocked PGE2-induced HCC cell growth and invasive ability in human HCC Huh-7 cells. The effect of PGE2 on c-
Myc expression was mainly through the EP4 receptor, and EP4 receptor-mediated c-Myc protein upregulation largely
depended on de novo biosynthesis of c-Myc mRNA and its protein. EP4 receptor signaling activated GS/AC and
increased the intracellular cAMP level in Huh-7 cells. The adenylate cyclase (AC) activator forskolin mimicked the effects
of the EP4 receptor agonist on c-Myc expression, while the AC inhibitor SQ22536 reduced EP4 receptor-mediated c-Myc
upregulation. These data confirm the involvement of the GS/AC/cAMP pathway in EP4 receptor-mediated c-Myc
upregulation. Moreover, the phosphorylation levels of CREB protein were markedly elevated by EP4 receptor signaling,
and by using specific inhibitor and siRNA interference, we demonstrated that PKA/CREB was also involved in the EP4
receptor-mediated c-Myc upregulation. In summary, the present study revealed that PGE2 significantly upregulates c-
Myc expression at both mRNA and protein levels through the EP4R/GS/AC/cAMP/PKA/CREB signaling pathway, thus
promoting cell growth and invasion in HCC cells. Targeting of the PGE2/EP4R/c-Myc pathway may be a new therapeutic
strategy to prevent and cure human HCC.
~3 Citings
83. TD-19, an erlotinib derivative, induces epidermal growth factor receptor wild-type nonsmall-cell lung cancer
apoptosis through CIP2A-mediated pathway
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By Chao, Ting-Ting; Wang, Cheng-Yi; Lai, Chih-Cheng; Chen, Yen-Lin; Tsai, Yi-Ting; Chen, Pao-Tzu; Lin, Hen-I.;
Huang, Yuh-Chin T.; Shiau, Chung-Wai; Yu, Chong-Jen; et al
From Journal of Pharmacology and Experimental Therapeutics (2014), 351(2), 352-358, 7 pp.. Language: English,
Database: CAPLUS, DOI:10.1124/jpet.114.215418
Some patients with nonsmall-cell lung cancer (NSCLC) without epidermal growth factor receptor (EGFR) mutations still
respond to gefitinib and erlotinib, suggesting that there may be a mechanism(s) other than the EGFR pathway that
mediates the tumoricidal effects. In the current study, we tested the efficacy of TD-19, a novel compd. chem. modified
from erlotinib, which has more potent apoptotic effects than erlotinib in EGFR wild-type NSCLC cell lines. TD-19 induced
significant cell death and apoptosis in H358, H441, H460, and A549 cells, as evidenced by increased caspase-3 activity
and cleavage of procaspase-9 and poly (ADP-ribose) polymerase. The apoptotic effect of TD-19 in H460 cells, which
were resistant to erlotinib, was assocd. with downregulation of cancerous inhibitor of protein phosphatase 2A (CIP2A),
increased protein phosphatase 2A (PP2A) activity, and decreased AKT phosphorylation, but minimal effects on EGFR
phosphorylation. Overexpression of CIP2A partially protected the H460 cells from TD-19-induced apoptosis. Okadaic
acid, a known PP2A inhibitor, significantly reduced TD-19-induced apoptosis, while forskolin, which increased PP2A
activity, increased the apoptotic effect of TD-19. TD-19 inhibited the growth of H460 xenograft tumors by ∼80%. We
conclude that TD-19 exerted tumoricidal effects on NSCLC cells. TD-19 provides proof that the CIP2A pathway may be
a novel target for the treatment of EGFR wild-type NSCLC.
~0 Citings
84. Studying chemical-regulation of intracellular kinase activity by peptide microarray-based assay with gold
nanoparticle probes
By Li, Tao; Su, Min; Ma, Lan; Liu, Dianjun; Wang, Zhenxin
From Analytical Methods (2014), 6(23), 9404-9409. Language: English, Database: CAPLUS,
DOI:10.1039/C4AY02157F
In the present work, the chem. regulation of intracellular kinase activity has been studied by a peptide microarray-based
resonance light scattering (RLS) assay with gold nanoparticle (AuNP) probes. After interactions of five cell lines and
seven compds. (six potential inhibitors of cyclic adenosine monophosphate (cAMP)-dependent protein kinase A (PKA)
and Forskolin (Fsk)), we found that the intracellular PKA activity is strongly regulated by the extracellular compds. and
that the compds. affect the intracellular PKA activity in a dose-dependent manner. The exptl. results demonstrate that
the peptide microarray-based RLS assay can be employed to quant. det. the efficiency of the inhibitor/activator on the
kinase activity at the cellular level. In addn., chem.-mediated fluctuation of the PKA activity in different cell phases (e.g.,
G2/M phase and M/G1 phase) was successfully detected, which also demonstrates the utility of the approach for
monitoring chem. regulation of intracellular kinase activity.
~0 Citings
85. Synthesis of cyclic 1,9-acetal derivatives of forskolin and their bioactivity evaluation
By Ponnam, Devendar; Shilpi, Singh; Srinivas, K. V. N. S.; Suiab, Luqman; Alam, Sarfaraz; Amtul, Zehra; Arigari,
Niranjan Kumar; Jonnala, Kotesh Kumar; Siddiqui, Lubna; Dubey, Vijaya; et al
From European Journal of Medicinal Chemistry (2014), 87, 735-744. Language: English, Database: CAPLUS,
DOI:10.1016/j.ejmech.2014.10.013
A new series of 1,9-acetals of forskolin, I (R1 = H, R2 = COMe, R3 = PH, 2-FC6H4, 2-furyl, etc.; R1 = R2 = H, R3 = Ph, Me;
R1 = COMe, R2 = H, R3 = Ph, Me), were synthesized by treating with arom. and aliph. aldehydes using Ceric ammonium
nitrate as catalyst and evaluated for anticancer and α-glucosidase inhibition activities. Among the synthesized compds., I
(R1 = R2 = H, R3 = Ph, Me; R1 = COMe, R2 = H, R3 = Ph) showed potential cytotoxic activity towards human cancer cell
lines MCF-7 (Human Breast Adenocarcinoma), MDA-MB (Human Breast Carcinoma), HeLa (Human Cervix
Adenocarcinoma), A498 (Human Kidney Carcinoma), K562 (Human Erythromyeloblastoid leukemia), SH-SY5Y (Human
Neuroblastoma), Hek293 (Human Embryonic Kidney) and WRL68 (Human Hepatic) with IC50 values ranging between
0.95 and 47.96 µg/mL. Osmotic fragility test revealed compd. I (R1 = COMe, R2 = H, R3 = Ph) as non-toxic to human
erythrocytes at the tested concns. of 50 and 100 µg/mL. Compds. I (R1 = H, R2 = COMe, R3 = 4-O2NC6H4) (IC50 value
0.76 µg/mL) and I (R1 = H, R2 = COMe, R3 = trans-CH:CHPh) (IC50 value 0.74 µg/mL) significantly inhibited α-
glucosidase in an in vitro system. In silico based docking, ADME and toxicity risk assessment studies also showed
discernible α-glucosidase activity for compds. I (R1 = H, R2 = COMe, R3 = 4-O2NC6H4, trans-CH:CHPh) compared to std.
acarbose.
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~4 Citings
86. High-throughput screen for pharmacoperones of the vasopressin type 2 receptor

By Conn, P. Michael; Smith, Emery; Hodder, Peter; Janovick, Jo Ann; Smithson, David C.
From Journal of Biomolecular Screening (2013), 18(8), 930-937, 8 pp.. Language: English, Database: CAPLUS,
DOI:10.1177/1087057113483559
Pharmacoperone drugs correct the folding of misfolded protein mutants and restore function (i.e., "rescue") by correcting
the routing of (otherwise) misrouted mutants. Assays for pharmacoperones have not been applied to screen large
libraries previously. Currently, most pharmacoperones possess intrinsic agonist or antagonist activities since these were
identified using high-throughput screens aimed at discovering direct agonists or antagonists. Here we describe an ultra-
high-throughput compatible no-wash assay system designed to specifically identify pharmacoperones of the vasopressin
type 2 receptor (V2R). Development of such assays is important and novel since useful chem. structures with the ability
to control cellular trafficking but lacking intrinsic agonist or antagonist properties have not likely been identified using
existing screens. In the described assay, the level of functional human V2R (hV2R) (mutant) present in each test well is
quantitated by stimulation with satg. levels of agonist followed by use of a luminescent-based cyclic adenosine
monophosphate assay. This allows the assay to identify compds. that increase the trafficking of mutant hV2R[L83Q] in
our model system.
~0 Citings
87. The river blindness drug Ivermectin and related macrocyclic lactones inhibit WNT-TCF pathway responses in
human cancer
By Melotti, Alice; Mas, Christophe; Kuciak, Monika; Lorente-Trigos, Aiala; Borges, Isabel; Ruiz i Altaba, Ariel
From EMBO Molecular Medicine (2014), 6(10), 1263-1278. Language: English, Database: CAPLUS,
DOI:10.15252/emmm.201404084
Constitutive activation of canonical WNT-TCF signaling is implicated in multiple diseases, including intestine and lung
cancers, but there are no WNT-TCF antagonists in clin. use. We have performed a repositioning screen for WNT-TCF
response blockers aiming to recapitulate the genetic blockade afforded by dominant-neg. TCF. We report that Ivermectin
inhibits the expression of WNT-TCF targets, mimicking dnTCF, and that its low concn. effects are rescued by direct
activation by TCFVP16. Ivermectin inhibits the proliferation and increases apoptosis of various human cancer types. It
represses the levels of C-terminal β-CATENIN phosphoforms and of CYCLIN D1 in an okadaic acid-sensitive manner,
indicating its action involves protein phosphatases. In vivo, Ivermectin selectively inhibits TCF-dependent, but not TCF-
independent, xenograft growth without obvious side effects. Anal. of single semi-synthetic derivs. highlights Selamectin,
urging its clin. testing and the exploration of the macrocyclic lactone chem. space. Given that Ivermectin is a safe anti-
parasitic agent used by > 200 million people against river blindness, our results suggest its addnl. use as a therapeutic
WNT-TCF pathway response blocker to treat WNT-TCF-dependent diseases including multiple cancers.
~7 Citings

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88. Metformin protects cardiomyocyte from doxorubicin induced cytotoxicity through an AMP-activated protein
kinase dependent signaling pathway: an in vitro study
By Kobashigawa, Laura C.; Xu, Yan Chun; Padbury, James F.; Tseng, Yi-Tang; Yano, Naohiro
From PLoS One (2014), 9(8), e104888/1-e104888/12, 12 pp.. Language: English, Database: CAPLUS,
Doxorubicin (Dox) is one of the most widely used antitumor drugs, but its cumulative cardiotoxicity have been major
concerns in cancer therapeutic practice for decades. Recent studies established that metformin (Met), an oral anti-
diabetic drug, provides protective effects in Dox-induced cardiotoxicity. Met has been shown to increase fatty acid
oxidn., an effect mediated by AMP activated protein kinase (AMPK). Here we delineate the intracellular signaling factors
involved in Met mediated protection against Dox-induced cardiotoxicity in the H9c2 cardiomyoblast cell line. Treatment
with low dose Met (0.1 mM) increased cell viabilities and Ki-67 expressions while decreasing LDH leakages, ROS
generations and [Ca2+]i. The protective effect was reversed by a co-treatment with compd.-C, an AMPK specific
inhibitor, or by an over expression of a dominant-neg. AMPKα cDNA. Inhibition of PKA with H89 or a suppression of Src
kinase by a small hairpin siRNA also abrogated the protective effect of the low dose Met. Whereas, with a higher dose of
Met (1.0 mM), the protective effects were abolished regardless of the enhanced AMPK, PKA/CREB1 and Src kinase
activity. In high dose Met treated cells, expression of platelet-derived growth factor receptor (PDGFR) was significantly
suppressed. Furthermore, the protective effect of low dose Met was totally reversed by co-treatment with AG1296, a
PDGFR specific antagonist. These data provide in vitro evidence supporting a signaling cascade by which low dose Met
exerts protective effects against Dox via sequential involvement of AMPK, PKA/CREB1, Src and PDGFR. Whereas high
dose Met reverses the effect by suppressing PDGFR expression.
~2 Citings
89. Searching for combinations of small-molecule correctors to restore F508del-cystic fibrosis transmembrane
conductance regulator function and processing
By Boinot, Clement; Souchet, Mathilde Jollivet; Ferru-Clement, Romain; Becq, Frederic
From Journal of Pharmacology and Experimental Therapeutics (2014), 350(3), 624-634, 11 pp.. Language: English,
Database: CAPLUS, DOI:10.1124/jpet.114.214890
The mutated protein F508del-cystic fibrosis transmembrane conductance regulator (CFTR) failed to traffic properly as a
result of its retention in the endoplasmic reticulum and functions as a chloride (Cl-) channel with abnormal gating and
endocytosis. Small chems. (called correctors) individually restore F508del-CFTR trafficking and Cl- transport function,
but recent findings indicate that synergistic pharmacol. should be considered to address CFTR defects more clearly. We
studied the function and maturation of F508del-CFTR expressed in HeLa cells using a combination of five correctors
[miglustat, IsoLAB (1,4-dideoxy-2-hydroxymethyl-1,4-imino-l-threitol), Corr4a (N-[2-(5-chloro-2-methoxy-phenylamino)-4'-
methyl-[4,5']bithiazolyl-2'-yl]-benzamide), VX-809 [3-(6-(1-(2,2-difluorobenzo[d][1,3]dioxol-5-
yl)cyclopropanecarboxamido)-3-methylpyridin-2-yl)benzoic acid], and suberoylamilide hydroxamic acid (SAHA)]. Using
the whole-cell patch-clamp technique, the c.d. recorded in response to CFTR activators (forskolin + genistein) was
significantly increased in the presence of the following combinations: VX-809 + IsoLAB; VX-809 + miglustat + SAHA; VX-
809 + miglustat + IsoLAB; VX-809 + IsoLAB + SAHA; VX-809 + miglustat + IsoLAB + SAHA. These combinations
restored the activity of F508del-CFTR but with a differential effect on the appearance of mature c-band of F508del-CFTR
proteins. Focusing on the VX-809 + IsoLAB cocktail, we recorded a level of correction higher at 37°C vs. room temp.,
but without amelioration of the thermal instability of CFTR. The level of functional rescue with VX-809 + IsoLAB after 4 h
of incubation was maximal and similar to that obtained in optimal conditions of use for each compd. (i.e., 24 h for VX-809
+ 4 h for IsoLAB). Finally, we compared the stimulation of F508del-CFTR by forskolin or forskolin + VX-770 [N-(2,4-di-
tert-butyl-5-hydroxyphenyl)-4-oxo-1,4-dihydroquinoline-3-carboxamide] with cells cor. by VX-809 + IsoLAB. Our results
open new perspectives for the development of a synergistic polypharmacol. to rescue F508del-CFTR and show the
importance of temp. on the effect of correctors and on the level of correction, suggesting that optimized combination of
correctors could lead to a better rescue of F508del-CFTR function.
~2 Citings
90. Metallothionein-3 (MT-3) in the Human Adrenal Cortex and its Disorders
By Felizola, Saulo J. A.; Nakamura, Yasuhiro; Arata, Yuki; Ise, Kazue; Satoh, Fumitoshi; Rainey, William E.;
Midorikawa, Sanae; Suzuki, Shinichi; Sasano, Hironobu
From Endocrine Pathology (2014), 25(3), 229-235. Language: English, Database: CAPLUS, DOI:10.1007/s12022-013-
9280-9
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Metallothionein-3 (MT-3) is an intracellular, low mol. wt. protein mainly distributed in the central nervous system but also
in various peripheral organs and several types of human neoplasms. However, details of MT-3 expression have not
been examd. in human adrenal cortex and its disorders. The mRNA levels of MT-3 were first evaluated by quant. RT-
PCR (qPCR) in adrenocortical aldosterone-producing adenoma (APA: 11) and cortisol-producing adenoma (CPA: 14). In
addn., MT-3 immunohistochem. was performed in non-pathol. adrenal glands (NA: 19), idiopathic hyperaldosteronism
(IHA: 10), APA (20), CPA (24), adjacent non-neoplastic adrenal glands of adenoma (AAG: 20), and adrenocortical
carcinoma (ACC: 8). H295R cells were also treated with angiotensin-II or forskolin in a time-dependent manner, and the
changes of MT-3 mRNA levels were evaluated by qPCR. Results of qPCR anal. demonstrated that MT-3 mRNA levels
were significantly higher in APA than CPA (P = 0.0004). MT-3 immunoreactivity was detected in the zona glomerulosa of
NA, IHA, and AAG, as well as in APA, CPA, and ACC. When treated with angiotensin-II and forskolin, MT-3 mRNA
levels reached a peak by 12 h in H295R cells, with significantly higher levels compared to control non-treated cells (P <
0.01). The presence of MT-3 in the ZG of NA, IHA, and AAG, as well as APA may imply a role in the pathophysiol. of
aldosterone-producing tissues.
~10 Citings
91. Steroidogenic enzymes, their related transcription factors and nuclear receptors in human sebaceous glands
under normal and pathological conditions
By Azmahani, Abdullah; Nakamura, Yasuhiro; Felizola, Saulo J. A.; Ozawa, Yohei; Ise, Kazue; Inoue, Takayoshi;
McNamara, Keely M.; Doi, Masao; Okamura, Hitoshi; Zouboulis, Christos C.; et al
From Journal of Steroid Biochemistry and Molecular Biology (2014), 144(Part_B), 268-279. Language: English,
Database: CAPLUS, DOI:10.1016/j.jsbmb.2014.07.010
The sebaceous gland is a major site of steroid synthesis in human skin, but details of the status of steroidogenic
enzymes and their regulation in human sebaceous glands under normal and pathol. conditions have rarely been
reported. Therefore, in this study, we examd. the status of steroidogenic enzymes, sex steroid receptors and
transcription factors in human sebaceous glands under normal and pathol. conditions to explore their possible roles in in
situ steroid prodn. in human skin. Immunohistochem. anal. was performed in a total of 59 human skin specimens,
including 22 normal human sebaceous glands, 12 with sebaceous nevus, 12 with sebaceous gland hyperplasia, 3 with
sebaceoma and 10 with sebaceous carcinoma. Immortalised human SZ95 sebocytes were treated with forskolin or
vehicle for 3 h, 6 h, 12 h or 24 h, and the mRNA levels of steroidogenic enzymes were evaluated at each time point using
quant. RT-PCR (qPCR). The results of immunohistochem. demonstrated the immunoreactivity of 3β-HSD1, CYP11A1,
StAR, 17β-HSD5, CYP17A1, 5α-red1, PRB, AR and NGFI-B in normal human sebaceous gland, with lower levels of
expression in pathol. sebaceous glands. The results of the in vitro study also indicated that the expression levels of 3β-
HSD1, CYP11A1, StAR, 5α-red1 and NGFI-B were elevated by forskolin. 3β-HSD1 and other steroidogenic enzymes
were expressed in sebaceous glands resulting in in situ androgen and progesterone synthesis and their functions.
~5 Citings
92. Quantitative impedimetric NPY-receptor activation monitoring and signal pathway profiling in living cells
By te Kamp, Verena; Lindner, Ricco; Jahnke, Heinz-Georg; Krinke, Dana; Kostelnik, Katja B.; Beck-Sickinger, Annette
G.; Robitzki, Andrea A.
From Biosensors & Bioelectronics (2015), 67, 386-393. Language: English, Database: CAPLUS,
DOI:10.1016/j.bios.2014.08.066
Label-free and non-invasive monitoring of receptor activation and identification of the involved signal pathways in living
cells is an ongoing analytic challenge and a great opportunity for biosensoric systems. In this context, we developed an
impedance spectroscopy-based system for the activation monitoring of NPY-receptors in living cells. Using an optimized
interdigital electrode array for sensitive detection of cellular alterations, we were able for the first time to quant. detect the
NPY-receptor activation directly without a secondary or enhancer reaction like cAMP-stimulation by forskolin. More
strikingly, we could show that the impedimetric based NPY-receptor activation monitoring is not restricted to the Y1-
receptor but also possible for the Y2- and Y5-receptor. Furthermore, we could monitor the NPY-receptor activation in
different cell lines that natively express NPY-receptors and proof the specificity of the obsd. impedimetric effect by
agonist/antagonist studies in recombinant NPY-receptor expressing cell lines. To clarify the nature of the obsd.
impedimetric effect we performed an equiv. circuit anal. as well as analyzed the role of cell morphol. and receptor
internalization. Finally, an antagonist based extensive mol. signal pathway anal. revealed small alterations of the actin
cytoskeleton as well as the inhibition of at least L-type calcium channels as major reasons for the obsd. NPY-induced
impedance increase. Taken together, our novel impedance spectroscopy based NPY-receptor activation monitoring
system offers the opportunity to identify signal pathways as well as for novel versatile agonist/antagonist screening
systems for identification of novel therapeutics in the field of obesity and cancer.
~2 Citings
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93. Quantification of dynamic morphological drug responses in 3D organotypic cell cultures by automated
image analysis
By Harma, Ville; Schukov, Hannu-Pekka; Happonen, Antti; Ahonen, Ilmari; Virtanen, Johannes; Siitari, Harri; Akerfelt,
Malin; Lotjonen, Jyrki; Nees, Matthias
Glandular epithelial cells differentiate into complex multicellular or acinar structures, when embedded in three-
dimensional (3D) extracellular matrix. The spectrum of different multicellular morphologies formed in 3D is a sensitive
indicator for the differentiation potential of normal, non-transformed cells compared to different stages of malignant
progression. In addn., single cells or cell aggregates may actively invade the matrix, utilizing epithelial, mesenchymal or
mixed modes of motility. Dynamic phenotypic changes involved in 3D tumor cell invasion are sensitive to specific small-
mol. inhibitors that target the actin cytoskeleton. We have used a panel of inhibitors to demonstrate the power of
automated image anal. as a phenotypic or morphometric readout in cell-based assays. We introduce a streamlined
stand-alone software soln. that supports large-scale high-content screens, based on complex and organotypic cultures.
AMIDA (Automated Morphometric Image Data Anal.) allows quant. measurements of large nos. of images and
structures, with a multitude of different spheroid shapes, sizes and textures. AMIDA supports an automated workflow
and can be combined with quality control and statistical tools for data interpretation and visualization. We have used a
representative panel of 12 prostate and breast cancer lines that display a broad spectrum of different spheroid
morphologies and modes of invasion, challenged by a library of 19 direct or indirect modulators of the actin cytoskeleton
which induce systematic changes in spheroid morphol. and differentiation vs. invasion. These results were
independently validated by 2D proliferation, apoptosis and cell motility assays. We identified three drugs that primarily
attenuated the invasion and formation of invasive processes in 3D, without affecting proliferation or apoptosis. Two of
these compds. block Rac signalling, one affects cellular cAMP/cGMP accumulation. Our approach supports the growing
needs for user-friendly, straightforward solns. that facilitate large-scale, cell-based 3D assays in basic research, drug
discovery and target validation.
~2 Citings
94. Ras Oncoprotein Disrupts the TSH/CREB Signaling Upstream Adenylyl Cyclase in Human Thyroid Cell
By Salzano, Marcella; Russo, Eleonora; Salzano, Salvatore; Bifulco, Maurizio; Vitale, Mario
DOI:10.1002/jcp.24672
Activating mutations in RAS genes and p21 Ras overactivation are common occurrences in a variety of human tumors.
P21 Ras oncoproteins deregulate a no. of signaling pathways, dedifferentiating the thyroid cell, and neg. regulating the
expression of thyroid specific genes. In rat thyroid cells, Ras oncoproteins inhibit the TSH pathway by reducing PKA
activity and thus the expression of thyroid specific genes, while in mouse melanocytes, Ras oncoproteins reduce the
αMSH-stimulated cAMP signaling by increasing the expression of the phosphodiesterase-4B. Given these cell-
dependent differences, we investigated if and how the TSH/CREB pathway is modulated by Ras oncoprotein in a human
thyroid cell line. CREB phosphorylation was stimulated by TSH and forskolin in TAD-2 cells. RasV12 expression neg.
regulated the TSH-stimulated CREB phosphorylation but was ineffective on forskolin-stimulated CREB phosphorylation.
Phosphodiesterase inhibition by IBMX enhanced TSH-stimulated CREB phosphorylation, but did not restore TSH-
stimulated CREB phosphorylation inhibited by Ras oncoprotein. These data indicate that Ras oncoprotein disrupts the
TSH/CREB pathway, upstream adenylyl cyclase, and highlight the existence of mechanisms of interaction between Ras
and the cAMP pathway different in human and in rat thyroid cells.
~0 Citings
95. A high throughput screening assay system for the identification of small molecule inhibitors of gsp
By Bhattacharyya, Nisan; Hu, Xin; Chen, Catherine Z.; Griner, Lesley A. Mathews; Zheng, Wei; Inglese, James; Austin,
Christopher P.; Marugan, Juan J.; Southall, Noel; Neumann, Susanne; et al
SciFinder® Page 39
Mis-sense mutations in the α-subunit of the G-protein, Gsα, cause fibrous dysplasia of bone/McCune-Albright syndrome.
The biochem. outcome of these mutations is constitutively active Gsα and increased levels of cAMP. The aim of this
study was to develop an assay system that would allow the identification of small mol. inhibitors specific for the mutant
Gsα protein, the so-called gsp oncogene. Com. available Chinese hamster ovary cells were stably transfected with
either wild-type (WT) or mutant Gsα proteins (R201C and R201H). Stable cell lines with equiv. transfected Gsα protein
expression that had relatively lower (WT) or higher (R201C and R201H) cAMP levels were generated. These cell lines
were used to develop a fluorescence resonance energy transfer (FRET)-based cAMP assay in 1536-well microplate
format for high throughput screening of small mol. libraries. A small mol. library of 343,768 compds. was screened to
identify modulators of gsp activity. A total of 1,356 compds. with inhibitory activity were initially identified and reconfirmed
when tested in concn. dose responses. Six hundred eighty-six mols. were selected for further anal. after removing
cytotoxic compds. and those that were active in forskolin-induced WT cells. These mols. were grouped by potency,
efficacy, and structural similarities to yield 22 clusters with more than 5 of structurally similar members and 144 singleton
mols. Seven chemotypes of the major clusters were identified for further testing and analyses.
~0 Citings
96. Improvement of chloride transport defect by gonadotropin-releasing hormone (GnRH) in cystic fibrosis
epithelial cells
By Benz, Nathalie; Le Hir, Sophie; Norez, Caroline; Kerbiriou, Mathieu; Calvez, Marie-Laure; Becq, Frederic; Trouve,
Pascal; Ferec, Claude
A review. Cystic fibrosis (CF), the most common autosomal recessive disease in Caucasians, is due to mutations in the
CFTR gene. F508del, the most frequent mutation in patients, impairs CFTR protein folding and biosynthesis. The
F508del-CFTR protein is retained in the endoplasmic reticulum (ER) and its traffic to the plasma membrane is altered.
Nevertheless, if it reaches the cell surface, it exhibits a Cl- channel function despite a short half-life. Pharmacol.
treatments may target the F508del-CFTR defect directly by binding to the mutant protein or indirectly by altering cellular
proteostasis, and promote its plasma membrane targeting and stability. We previously showed that annexine A5
(AnxA5) directly binds to F508del-CFTR and, when overexpressed, promotes its membrane stability, leading to the
restoration of some Cl- channel function in cells. Because Gonadotropin-Releasing Hormone (GnRH) increases AnxA5
expression in some cells, we tested it in CF cells. We showed that human epithelial cells express GnRH-receptors
(GnRH-R) and that GnRH induces an AnxA5 overexpression and an increased Cl- channel function in F508del-CFTR
cells, due to an increased stability of the protein in the membranes. Beside the numerous physiol. implications of the
GnRH-R expression in epithelial cells, we propose that a topical use of GnRH is a potential treatment in CF.
~1 Citing
97. Stress-related hormone norepinephrine induces interleukin-6 expression in GES-1 cells

By Yang, R.; Lin, Q.; Gao, H. B.; Zhang, P.
From Brazilian Journal of Medical and Biological Research (2014), 47(2), 101-109, 9. Language: English, Database:
CAPLUS, DOI:10.1590/1414-431X20133346
In the current literature, there is evidence that psychol. factors can affect the incidence and progression of some cancers.
Interleukin 6 (IL-6) is known to be elevated in individuals experiencing chronic stress and is also involved in oncogenesis
and cancer progression. However, the precise mechanism of IL-6 induction by the stress-related hormone
norepinephrine (NE) is not clear, and, furthermore, there are no reports about the effect of NE on IL-6 expression in
gastric epithelial cells. In this study, we examd. the effect of NE on IL-6 expression in immortalized human gastric
epithelial cells (GES-1 cells). Using realtime PCR and enzyme-linked immunoassay, we demonstrated that NE can
induce IL-6 mRNA and protein expression in GES-1 cells. The induction is through the β-adrenergic receptor-cAMP-
protein kinase A pathway and mainly at the transcriptional level. Progressive 5'-deletions and site-directed mutagenesis
of the parental construct show that, although activating-protein-1 (AP-1), cAMP-responsive element binding protein
(CREB), CCAAT-enhancer binding protein-β (C/EBP-β), and nuclear factor κ-light-chain-enhancer of activated B cells
(NF-κB) binding sites are all required in the basal transcription of IL-6, only AP-1 and CREB binding sites in the IL-6
promoter are required in NE-induced IL-6 expression. The results suggest that chronic stress may increase IL-6
secretion of human gastric epithelial cells, at least in part, by the stress-assocd. hormone norepinephrine, and provides
basic data on stress and gastric cancer progression.
~5 Citings

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98. Hyperphosphorylation of PP2A in colorectal cancer and the potential therapeutic value showed by its
forskolin-induced dephosphorylation and activation
By Cristobal, Ion; Rincon, Raul; Manso, Rebeca; Madoz-Gurpide, Juan; Carames, Cristina; del Puerto-Nevado, Laura;
Rojo, Federico; Garcia-Foncillas, Jesus
From Biochimica et Biophysica Acta, Molecular Basis of Disease (2014), 1842(9), 1823-1829. Language: English,
Database: CAPLUS, DOI:10.1016/j.bbadis.2014.06.032
The tumor suppressor protein phosphatase 2A (PP2A) is frequently inactivated in human cancer and phosphorylation of
its catalytic subunit (p-PP2A-C) at tyrosine-307 (Y307) has been described to inhibit this phosphatase. However, its mol.
and clin. relevance in colorectal cancer (CRC) remains unclear. P-PP2A-C Y307 was detd. by immunoblotting in 7 CRC
cell lines and 35 CRC patients. CRC cells were treated with the PP2A activator forskolin alone or combined with the
PP2A inhibitor okadaic acid, 5-fluorouracil and oxaliplatin. We examd. cell growth, colonosphere formation, caspase
activity and AKT and ERK activation. PP2A-C was found hyperphosphorylated in CRC cell lines. Forskolin
dephosphorylated and activated PP2A, impairing proliferation and colonosphere formation, and inducing activation of
caspase 3/7 and changes in AKT and ERK phosphorylation. Moreover, forskolin showed additive effects with 5-
fluorouracil and oxaliplatin treatments. Anal. of p-PP2A-C Y307 in primary tumors confirmed the presence of this
alteration in a subgroup of CRC patients. Our data show that PP2A-C hyperphosphorylation is a frequent event that
contributes to PP2A inhibition in CRC. Antitumoral effects of forskolin-mediated PP2A activation suggest that the anal. of
p-PP2A-C Y307 status could be used to identify a subgroup of patients who would benefit from treatments based on
PP2A activators.
~4 Citings
99. Ultraviolet radiation, aging and the skin: prevention of damage by topical cAMP manipulation
By Amaro-Ortiz, Alexandra; Yan, Betty; D'Orazio, John A.
From Molecules (2014), 19(5), 6202-6219, 18 pp.. Language: English, Database: CAPLUS,
DOI:10.3390/molecules19056202
A review. Being the largest and most visible organ of the body and heavily influenced by environmental factors, skin is
ideal to study the long-term effects of aging. Throughout our lifetime, we accumulate damage generated by UV radiation.
UV causes inflammation, immune changes, phys. changes, impaired wound healing and DNA damage that promotes
cellular senescence and carcinogenesis. Melanoma is the deadliest form of skin cancer and among the malignancies of
highest increasing incidence over the last several decades. Melanoma incidence is directly related to age, with highest
rates in individuals over the age of 55 years, making it a clear age-related disease. In this review, we will focus on UV-
induced carcinogenesis and photo aging along with natural protective mechanisms that reduce amt. of "realized" solar
radiation dose and UV-induced injury. We will focus on the theor. use of forskolin, a plant-derived pharmacol. active
compd. to protect the skin against UV injury and prevent aging symptoms by up-regulating melanin prodn. We will
discuss its use as a topically-applied root-derived formulation of the Plectranthus barbatus (Coleus forskolii) plant that
grows naturally in Asia and that has long been used in various Aryuvedic teas and therapeutic prepns.
~4 Citings
100. Ghrelin and des-acyl ghrelin inhibit aromatase expression and activity in human adipose stromal cells:
suppression of cAMP as a possible mechanism
By Docanto, Maria M.; Yang, Fangyuan; Callaghan, Brid; Au, CheukMan C.; Ragavan, Rahini; Wang, Xuyi; Furness,
John B.; Andrews, Zane B.; Brown, Kristy A.
DOI:10.1007/s10549-014-3060-1
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Aromatase converts androgens into estrogens and its expression within adipose stromal cells (ASCs) is believed to be
the major driver of estrogen-dependent cancers in older women. Ghrelin is a gut-hormone that is involved in the
regulation of appetite and known to bind to and activate the cognate ghrelin receptor, GHSR1a. The unacylated form of
ghrelin, des-acyl ghrelin, binds weakly to GHSR1a but has been shown to play an important role in regulating a no. of
physiol. processes. The aim of this study was to det. the effect of ghrelin and des-acyl ghrelin on aromatase in primary
human ASCs. Primary human ASCs were isolated from adipose tissue of women undergoing cosmetic surgery. Real-
time PCR and tritiated water-release assays were performed to examine the effect of treatment on aromatase transcript
expression and aromatase activity, resp. Treatments included ghrelin, des-acyl ghrelin, obestatin, and capromorelin
(GHSR1a agonist). GHSR1a protein expression was assessed by Western blot and effects of treatment on Ca2+ and
cAMP second messenger systems were examd. using the Flexstation assay and the Lance Ultra cAMP kit, resp. Results
demonstrate that pM concns. of ghrelin and des-acyl ghrelin inhibit aromatase transcript expression and activity in ASCs
under basal conditions and in PGE2-stimulated cells. Moreover, the effects of ghrelin and des-acyl ghrelin are mediated
via effects on aromatase promoter PII-specific transcripts. Neither the GHSR1a-specific agonist capromorelin nor
obestatin had any effect on aromatase transcript expression or activity. Moreover, GHSR1a protein was undetectable by
Western blot and neither ghrelin nor capromorelin elicited a calcium response in ASCs. Finally, ghrelin caused a
significant decrease in basal and forskolin-stimulated cAMP in ASC. These findings suggest that ghrelin acts at alternate
receptors in ASCs by decreasing intracellular cAMP levels. Ghrelin mimetics may be useful in the treatment of estrogen-
dependent breast cancer.
~4 Citings
101. Forskolin regulates bone sialoprotein gene expression in human breast cancer cells
By Kaneko, Hirotoshi; Kim, Dong-Soon; Ogata, Yorimasa
From International Journal of Oral-Medical Sciences (2013), 12(3), 196-203. Language: English, Database: CAPLUS,
DOI:10.5466/ijoms.12.196
Bone sialoprotein (BSP) is a prominent mineral-assocd. protein in the extracellular matrix of bone that has been
implicated in the metastatic activity of cancer cells. Forskolin (FSK) is a labdanediterpene that is produced by the Indian
Coleus plant. FSK elevates cAMP (cAMP) via adenylate cyclase activated protein kinase A (PKA), and it is widely used
to study cAMP regulation of cell processes. Here, it is reported how FSK regulates BSP gene transcription in human
breast cancer MCF7 cells. FSK (1 µM) increased BSP, Runx2 and Osterix mRNA levels in MCF7 cells at 12 h. From
transient transfection analyses using various BSP promoter-luciferase constructs, a cAMP response element (CRE), a
runt homodomain protein 2 (Runx2) and a fibroblast growth factor 2 (FGF2) response element (FRE) were identified as a
target of transcriptional activation by FSK. Gel mobility shift analyses showed that FSK increased binding of CRE and
FRE. These studies demonstrate that FSK stimulates BSP transcription in MCF7 cells by targeting the CRE and FRE
elements in the BSP gene promoter.
~0 Citings
102. In vitro culture studies on medicinal herb - Coleus forskohlii Briq

By Praveena, R.; Amarnath Pandian, S.; Jegadeesan, M.
From Libyan Agriculture Research Center Journal International (2012), 3(1), 30-35, 6 pp.. Language: English,
Database: CAPLUS
Coleus forskohlii Briq. (Lamiaceae) is an important medicinal plant with excellent export potential in herbal drug trade.
The root of this plant is medicinally useful for high blood pressure, spasmolysis, obesity and constipation. A diterpene
compd., forskolin obtained from tuberous root of this plant has been used for glaucoma congestive, asthma and certain
cancers. The present work was aimed at evolving a protocol for rapid multiplication using culture technique. Tissue
explants from leaf lamina, node and shoot tip were cultured on MS medium supplemented with different concns. (0.5-2.0
mg/l) of IBA, 2,4-D and BAP and their growth responses like callusing and shooting and rooting were elucidated. It was
obsd. that shoot tip elicited max. callusing, shooting and rooting response than that of leaf lamina and node. Multiple
shoots were obtained from leaf lamina explants. This indicates the possibility of using tissue culture protocol for com.
scale of prodn.
~0 Citings
103. Protective effect of an antithyroid compound against γ-radiation-induced damage in human colon cancer
cells
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By Perona, Marina; Dagrosa, Maria A.; Pagotto, Romina; Casal, Mariana; Pignataro, Omar; Pisarev, Mario A.; Juvenal,
Guillermo J.
From Radiation and Environmental Biophysics (2014), 53(3), 611-619. Language: English, Database: CAPLUS,
DOI:10.1007/s00411-014-0542-9
We have previously reported the radioprotective effect of propylthiouracil (PTU) on thyroid cells. The aim of the present
study was to analyze whether tumor cells and normal cells demonstrate the same response to PTU. Human colon
carcinoma cells were irradiated with γ-irradn. with or without PTU. We evaluated the clonogenic survival, intracellular
reactive oxygen species levels, catalase, superoxide dismutase and glutathione peroxidase activities, and apoptosis by
nuclear cell morphol. and caspase-3 activity assays. CAMP (cAMP) levels were measured by RIA. PTU treatment
increased surviving cell fraction at 2 Gy (SF2) from 56.9 ± 3.6 in controls to 75.0 ± 3.5 (p < 0.05) and diminished
radiation-induced apoptosis. In addn., we obsd. that the level of antioxidant enzymes' activity was increased in cells
treated with PTU. Moreover, pretreatment with PTU increased intracellular levels of cAMP. Forskolin (p < 0.01) and
dibutyryl cAMP (p < 0.05) mimicked the effect of PTU on SF2. Co-treatment with H89, an inhibitor of protein kinase A,
abolished the radioprotective effect of PTU. PTU reduces the toxicity of ionizing radiation by increasing cAMP levels and
also possibly through a redn. in apoptosis levels and in radiation-induced oxidative stress damage. We therefore
conclude that PTU protects both normal and cancer cells during exposure to radiation in conditions mimicking the
radiotherapy.
~0 Citings
104. Application of extract solution composition of natural multiple-form arsenic in producing the medical
formulations for resisting tumor stem cells, hepatitis B virus and resisting AIDS virus
By Zhang, Jinghong; Yang, Li
The ext. soln. compn. comprises natural multiple-form arsenic ext. soln. 0.001-99.9%, and addnl. at least one antitumor
agent. The antitumor agent is from camptothecin, catharanthine, vincristine, paclitaxel, tumor necrosis factor, interleukin,
interferon, transform growth factor-β, lipopolysaccharide, phorbol ester, adenylate cyclase activator an/or bone
morphogenetic protein 4. The natural multiple-form arsenic is from arsenic acid (ASIII), arsenious acid (AsV),
monomethyl arsenic acid (MMAIII), monomethyl arsenious acid (MMAV), di-Me arsenic acid (DMAIII), di-Me arsenious
acid (DMAV), monomethyl monothioarsenic acid (MMTA), di-Me dithioarsenic acid (DMDTA), tri-Me arsenic oxide
(TMAO), arsenobetaine (AsB), or arsenocholine (AsC). The ext. soln. compn. can be used in producing the medical
formulations (such as soft capsule, hard capsule, dripping pill, tablet, granule, injection, powder injection, oral soln.,
microcapsule, aerosol or suppository) for resisting tumor stem cells, hepatitis B virus and resisting AIDS virus, by oral
administration, injection or rectal administration, which expands the application range for the exploitation and utilization of
natural arsenical.
~0 Citings
105. Inhibition of CYP17A1 activity by resveratrol, piceatannol, and synthetic resveratrol analogs
By Oskarsson, Agneta; Spatafora, Carmela; Tringali, Corrado; Andersson, Aasa Ohlsson
From Prostate (Hoboken, NJ, United States) (2014), 74(8), 839-851. Language: English, Database: CAPLUS,
DOI:10.1002/pros.22801
BACKGROUND : Resveratrol (RSV) and resveratrol analogs have a potential use in prostate cancer chemoprevention
due to effects on for example, cell growth, apoptosis, angiogenesis, and metastasis. However, inhibition of CYP17A1, a
key enzyme in the androgen biosynthesis and a target for prostate cancer therapy, has not been explored as a possible
mechanism behind the effects on prostate cancer. METHODS : Human adrenocortical carcinoma cells, H295R, were
treated with RSV, piceatannol (PIC), 3,5,4'-triacetylresveratrol (RSVTA), 3,5-diacetylresveratrol (RSVDA), and 3,5,4'-
trimethylresveratrol (RSVTM) for 24 h at concns. of 1, 5, 10, 25, and 50 µM. Steroid secretion, enzyme activities, and
gene expression of key steps in steroidogenesis were investigated. RESULTS : Secretion of dihydroepiandrosterone
(DHEA), testosterone, and cortisol were drastically decreased by all test compds. at concns. that did not affect cell
viability. Progesterone and aldosterone secretion were increased. This steroid secretion pattern can be explained by the
demonstrated inhibition of CYP17A1 enzyme activity. The most efficient CYP17A1 inhibitors were the synthetic analogs
RSVTA, RSVDA, and RSVTM. Inhibition by RSVTM was more selective on the 17,20-lyase activity than hydroxylase
activity of CYP17A1. Treatment of cells with all compds., except RSVTM, caused increased estradiol levels, which could
be explained by the demonstrated inhibition of estrogen sulfate conjugation, catalyzed by SULT1E1. CONCLUSIONS :
Our results on CYP17A1 inhibition of RSV and RSV analogs suggest a novel mechanism for chemoprevention of
prostate cancer by resveratrol and the analogs. Esp. RSVTM, which has a preferential inhibition on the 17,20-lyase
activity of CYP17A1, may be a promising candidate for prostate cancer chemoprevention. Prostate 74:839-851, 2014. ©
2014 Wiley Periodicals, Inc.
~5 Citings
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106. Different effects of adenylyl cyclase activators and phosphodiesterases inhibitors on cervical cancer (HeLa)
and breast cancer (MCF-7) cells proliferation
By Mahdian, Davood; Shafiee-Nick, Reza; Mousavi, Seyed Hadi
From Toxicology Mechanisms and Methods (2014), 24(4), 307-314. Language: English, Database: CAPLUS,
DOI:10.3109/15376516.2014.898354
Breast and cervical cancers are the most common cancers in Iran and worldwide. Hormonal stimulation of cyclic
adenosine mono phosphate (cAMP) and the cAMP-dependent protein kinase PKA regulates cell growth by different
mechanism. cAMP can stimulate cell growth in many cell types while inhibiting cell growth in others. In some cell lines
have been shown that the proliferation of tumor cells is reduced by increasing cAMP in cells. In this study, we evaluate
growth arrest of selective PDE3 and non-selective PDE inhibitors, which lead to increase level of cAMP in cervical
(HeLa) and breast cancer (MCF7) cell lines have been studied. Cells were incubated with different concns. of selective,
non-selective PDE inhibitors, beta adrenergic receptor agonist and direct stimulator of adenylyl cyclase. Cell viability was
quantitated by MTT assay. Apoptotic cells were detd. using PI staining of DNA fragmentation by flow cytometry (sub-G1
peak). Result showed that selective PDE inhibitors decreased cell viability in HeLa and MCF-7 cells as a time-dependent
manner. Non-selective inhibitor and beta-adrenergic receptor agonist also decrease cell viability but they are less
powerful than selective PDE3 inhibitors. Forskolin had no effect in viability of cells. Anal. of DNA fragmentation by flow
cytometry showed apoptosis involved in selective PDE3 inhibitors induced toxicity in HeLa cell. Thus, the growth
inhibitory effects of selective PDE3 inhibitors are more effective than non-selective inhibitor. Further studies are needed
to investigate the mechanism of action is on the field.
~1 Citing
107. Preparation of triptolide derivatives with Cunninghamella

By Zhang, Jinghong; Xie, Shenxia; Tang, Yuanyuan
The invention provides a method for prepg. triptolide deriv., and its products and application thereof. The prepn. method
uses triptolide as microorganism metab. medicament, through special elicitor for microorganism metab. of triptolide,
simultaneously obtains multiple, high yield, triptolide-based hydroxylation and/or methylation modified triptolide deriv. as
shown in formula on Pg4, wherein R1, R2, R3 and R4 are independently H or alkyl, which may be substituted by alkoxy,
alkyl or hydroxy; group X has two hydroxy at most. The method includes (1) culturing preserved strain for 7 days,
blowing spores and prepg. spore suspension, (2) inoculating in potato liq. medium, oscillation culturing, adding elicitor,
culturing, inoculating triptolide, culturing till conversion rate of 0.5-3%, filtering, centrifugating filter liquor to obtain fermn.
extn. liquor, (3) filtering, extg. with Et acetate for 3 times, combining extn. liquor, extg. mycelia with Et acetate, filtering,
combining filter liquor with extn. liquor, concg., (4) dissolving residues in acetone, loading on silica gel column, eluting
with petroleum ether-Et acetate at a gradient concn., vaporizing eluates, dissolving in methanol, filtering to obtain sepn.
liquor, (5) purifying and crystg. to obtain triptolide derivs. The transforming strain is Cunninghamella blakesleana,
Cunninghamella echinulata, Cunninghamella elegans, Cunninghamella echinulata var. elegans, Cunninghamella
echinulata(Thaxter) Thaxter var. echinulata, Aspergillus niger, Aspergillus flavus, Mucor spinosus, Mucor subtilissimus,
Alternaria alternata, Penicillium janthinellum or Penicillium urticae. The elicitor is two or more of A, B and C, wherein A is
α-cyclodextrin, β-cyclodextrin, 2,6-dimethyl-β-cyclodextrin, etc.; B is Me jasmonate, salicylic acid, chitosan, etc.; C is
carbamazepine, modafinil, nevirapine, rifampicin, Hypericum perforatum, etc. These triptolide deriv. has good function of
inhibiting tumor, AIDS, and hepatitis B virus. The inventive method is environmentally friendly, has high selectivity, high
yield, can large-scale prep. triptolide deriv. using microorganism.
~1 Citing
108. Pharmacologic induction of epidermal melanin and protection against sunburn in a humanized mouse
model
By Ortiz, Alexandra Amaro; Vanover, Jillian C.; Scott, Timothy L.; Orazio, John A. D.
From Journal of Visualized Experiments (2013), (79), e50670/1-e50670/10. Language: English, Database: CAPLUS,
DOI:10.3791/50670
SciFinder® Page 44
Fairness of skin, UV sensitivity and skin cancer risk all correlate with the physiol. function of the melanocortin 1 receptor,
a Gs-coupled signaling protein found on the surface of melanocytes. Mc1r stimulates adenylyl cyclase and cAMP prodn.
which, in turn, up-regulates melanocytic prodn. of melanin in the skin. In order to study the mechanisms by which Mc1r
signaling protects the skin against UV injury, this study relies on a mouse model with "humanized skin" based on
epidermal expression of stem cell factor (Scf). K14-Scf transgenic mice retain melanocytes in the epidermis and
therefore have the ability to deposit melanin in the epidermis. In this animal model, wild type Mc1r status results in
robust deposition of black eumelanin pigment and a UV-protected phenotype. In contrast, K14-Scf animals with
defective Mc1r signaling ability exhibit a red/blonde pigmentation, very little eumelanin in the skin and a UV-sensitive
phenotype. Reasoning that eumelanin deposition might be enhanced by topical agents that mimic Mc1r signaling, we
found that direct application of forskolin ext. to the skin of Mc1r-defective fair-skinned mice resulted in robust eumelanin
induction and UV protection 1. Here we describe the method for prepg. and applying a forskolin-contg. natural root ext.
to K14-Scf fair-skinned mice and report a method for measuring UV sensitivity by detg. minimal erythematous dose
(MED). Using this animal model, it is possible to study how epidermal cAMP induction and melanization of the skin affect
physiol. responses to UV exposure.
~1 Citing
109. High-Content pSTAT3/1 Imaging Assays to Screen for Selective Inhibitors of STAT3 Pathway Activation in
Head and Neck Cancer Cell Lines
By Johnston, Paul A.; Sen, Malabika; Hua, Yun; Camarco, Daniel; Shun, Tong Ying; Lazo, John S.; Grandis, Jennifer
R.
From Assay and Drug Development Technologies (2014), 12(1), 55-79. Language: English, Database: CAPLUS,
DOI:10.1089/adt.2013.524
The oncogenic transcription factor signal transducer and activator of transcription 3 (STAT3) is hyperactivated in most
cancers and represents a plausible therapeutic target. In the absence of STAT3-selective small-mol. inhibitors, we
sought to develop pSTAT3/1 high-content imaging (HCS) assays to screen for selective inhibitors of STAT3 pathway
activation in head and neck squamous cell carcinomas (HNSCC) tumor cell lines. Based on the expression of the
interleukin-6 (IL-6)Rα and gp130 subunits of the IL-6 receptor complex and STAT3, we selected the Cal33 HNSCC cell
line as our model. After developing image acquisition and anal. procedures, we rigorously investigated the cytokine
activation responses to optimize the dynamic ranges of both assays and demonstrated that the pan-Janus kinase
inhibitor pyridone 6 nonselectively inhibited pSTAT3 and pSTAT1 activation with 50% inhibition concns. of 7.19±4.08 and
16.38±8.45 nM, resp. The optimized pSTAT3 HCS assay performed very well in a pilot screen of 1,726 compds. from
the Library of Pharmacol. Active Compds. and the National Institutes of Health clin. collection sets, and we identified 51
inhibitors of IL-6-induced pSTAT3 activation. However, only three of the primary HCS actives selectively inhibited STAT3
compared with STAT1. Our follow-up studies indicated that the nonselective inhibition of cytokine induced pSTAT3 and
pSTAT1 activation by G-alpha stimulatory subunit-coupled G-protein-coupled receptor agonists, and forskolin was likely
due to cyclic adenosine monophosphate-mediated up-regulation of suppressors of cytokine signaling 3. Azelastine, an
H1 receptor antagonist approved for the treatment of seasonal allergic rhinitis, nonallergic vasomotor rhinitis, and ocular
conjunctivitis, was subsequently confirmed as a selective inhibitor of IL-6-induced pSTAT3 activation that also reduced
the growth of HNSCC cell lines. These data illustrate the power of a chem. biol. approach to lead generation that utilizes
fully developed and optimized HCS assays as phenotypic screens to interrogate specific signaling pathways.
~7 Citings
110. Expression of mRNAs of urocortin in the STKM-1 gastric cancer cell line
By Akiyoshi, Kohei; Kamada, Minori; Fujioka, Kouki; Ikeda, Keiichi; Tojo, Katsuyoshi; Manome, Yoshinobu
From Anticancer Research (2013), 33(12), 5289-5294. Language: English, Database: CAPLUS
Background: Urocortin is analogous to corticotrophin-releasing factors (CRFs) and a member of the CRF family. We
previously demonstrated that urocortin mRNAs were expressed in both human and rat glioma cell lines, and that some of
these lines transcribed the receptors. We hypothesize that urocortin might also be expressed in a gastric cancer cell line.
The aim of the present study was to clarify the expression of mRNAs of urocortin1 (UCN1), -2 and -3 and of CRF and
CRF receptors 1 and 2 in a gastric cancer cell line. Materials and Methods: STKM-1 a poorly-differentiated
adenocarcinoma cell line was used. Transcripts in the cells were analyzed using cDNA. The fluctuation of mRNA with
cellular stress, such as the one caused by a chemotherapeutic agent, serum supplementation and forskolin was examd.
Results: Transcripts of UCN1, -2 and CRFR2 were expressed. No changes in transcription of UCN1 and UCN2 were
obsd. with cellular stress. However, expression of CRFR2 mRNA transcripts significantly increased after an initial 24-h
exposure to forskolin. Conclusion: Expression of the mRNAs of UCN1, 2 and CRFR2 was confirmed in the human
gastric cancer cell line, STKM-1. Although the quantity of CRFR2 transcripts varied with forskolin, the overall
transcription pattern was not influenced by cellular stimuli.
~1 Citing
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111. Methods of treating cancer using 3-(4-((4-(morpholinomethyl)benzyl)oxy)-1-oxoisoindolin-2-yl)piperidine-

2,6-dione
By Schafer, Peter H.; Gandhi, Anita
Provided herein are methods of treating, preventing and/or
managing cancers, which comprise administering to a patient
3-(4-((4-(morpholinomethyl)benzyl)oxy)-1-oxoisoindolin-2-yl)
piperidine-2,6-dione, or an enantiomer or a mixt. of
enantiomers thereof, or a pharmaceutically acceptable salt,
solvate, hydrate, co-crystal, clathrate, or polymorph thereof.
~0 Citings
112. Inactivation of the Carney complex gene 1 (PRKAR1A) alters spatiotemporal regulation of cAMP and cAMP-
dependent protein kinase: a study using genetically encoded FRET-based reporters
By Cazabat, Laure; Ragazzon, Bruno; Varin, Audrey; Potier-Cartereau, Marie; Vandier, Christophe; Vezzosi, Delphine;
Risk-Rabin, Marthe; Guellich, Aziz; Schittl, Julia; Lechene, Patrick; et al
From Human Molecular Genetics (2014), 23(5), 1163-1174. Language: English, Database: CAPLUS,
DOI:10.1093/hmg/ddt510
Carney complex (CNC) is a hereditary disease assocg. cardiac myxoma, spotty skin pigmentation and endocrine
overactivity. CNC is caused by inactivating mutations in the PRKAR1A gene encoding PKA type I alpha regulatory
subunit (RIα). Although PKA activity is enhanced in CNC, the mechanisms linking PKA dysregulation to endocrine
tumorigenesis are poorly understood. In this study, the authors used Foerster resonance energy transfer (FRET)-based
sensors for cAMP and PKA activity to define the role of RIα in the spatiotemporal organization of the cAMP/PKA
pathway. RIα knockdown in HEK293 cells increased basal as well as forskolin or prostaglandin E1 (PGE1)-stimulated
total cellular PKA activity as reported by western blots of endogenous PKA targets and the FRET-based global PKA
activity reporter, AKAR3. Using variants of AKAR3 targeted to subcellular compartments, the authors identified similar
increases in the response to PGE1 in the cytoplasm and at the outer mitochondrial membrane. In contrast, at the plasma
membrane, the response to PGE1 was decreased along with an increase in basal FRET ratio. These results were
confirmed by western blot anal. of basal and PGE1-induced phosphorylation of membrane-assocd. vasodilator-stimulated
phosphoprotein. Similar differences were obsd. between the cytoplasm and the plasma membrane in human adrenal
cells carrying a RIα inactivating mutation. RIα inactivation also increased cAMP in the cytoplasm, at the outer
mitochondrial membrane and at the plasma membrane, as reported by targeted versions of the cAMP indicator Epac1-
camps. These results show that RIα inactivation leads to multiple, compartment-specific alterations of the cAMP/PKA
pathway revealing new aspects of signaling dysregulation in tumorigenesis.
~4 Citings
113. cAMP-PKA inhibition of SK3 channel reduced both Ca2+ entry and cancer cell migration by regulation of
SK3-Orai1 complex
By Clarysse, Lucie; Gueguinou, Maxime; Potier-Cartereau, Marie; Vandecasteele, Gregoire; Bougnoux, Philippe;
Chevalier, Stephan; Chantome, Aurelie; Vandier, Christophe
From Pfluegers Archiv (2014), 466(10), 1921-1932. Language: English, Database: CAPLUS, DOI:10.1007/s00424-
013-1435-5
SciFinder® Page 46
SK3 channel mediates the migration of various cancer cells. When expressed in breast cancer cells, SK3 channel forms
a complex with Orai1, a voltage-independent Ca2+ channel. This SK3-Orai1 complex assocs. within lipid rafts where it
controls a constitutive Ca2+ entry leading to cancer cell migration and bone metastases development. Since cAMP was
found to modulate breast cancer cell migration, we hypothesized that this could be explained by a modulation of SK3
channel activity. Herein, we study the regulation of SK3 channel by the cAMP-PKA pathway and the consequences for
SK3-dependent Ca2+ entry and cancer cell migration. We established that the beta-adrenergic receptor agonist,
isoprenaline, or the direct adenylyl cyclase activator forskolin alone or in combination with the PDE4 inhibitor, CI-1044,
decreased SK3 channel activity without modifying the expression of SK3 protein at the plasma membrane. Forskolin and
CI-1044 reduced the SK3-dependent constitutive Ca2+ entry and the SK3-dependent migration of MDA-MB-435s cells.
PKA inhibition with KT 5720 reduced: (1) the effect of forskolin and CI-1044 by 50 % on Ca2+ entry and (2) SK3 activity
by inhibiting the serine phosphorylation of SK3. These cAMP-elevating agents displaced Orai1 protein outside lipid rafts
in contrast to SK3, which remained in the lipid rafts fractions. All together, these results show that activation of the
cAMP-PKA pathway decreases SK3 channel and SK3-Orai1 complex activities, leading to a decrease in both Ca2+ entry
and cancer cell migration. This work supports the potential use of cAMP-elevating agents to reduce cancer cell migration
and may provide novel opportunities to address/prevent bone metastasis.
~8 Citings
114. Cyclic AMP enhances progesterone action in human myometrial cells

By Chen, Li; Lei, Kaiyu; Malawana, Johann; Yulia, Angela; Sooranna, Suren R.; Bennett, Phillip R.; Liang, Zhiqing;
Grammatopoulos, Dimitri; Johnson, Mark R.
From Molecular and Cellular Endocrinology (2014), 382(1), 334-343. Language: English, Database: CAPLUS,
DOI:10.1016/j.mce.2013.10.018
CAMP has been shown to promote progesterone and glucocorticoid action in a variety of cellular settings. In this study,
we have used human myometrial cells to investigate whether cAMP potentiates the ability of progesterone to repress IL-
1β-driven COX-2 expression. We found that forskolin enhanced progesterone-repression of IL-1β-driven COX-2
expression in assocn. with delayed IL-1β-induced nuclear phospho-p65 entry and reduced NF-κB binding to the COX-2
promoter. Further, forskolin enhanced the progesterone-induced expression of FKBP5 and 11βHSD1, progesterone-
driven activity of a progesterone response element (PRE) and progesterone receptor (PR)-B binding to a transfected
PRE. In addn., forskolin treatment increased PR-B levels and reduced the PR-A:PR-B ratio while acutely decreasing the
assocn. between PR and nuclear receptor co-repressor (NCoR) and reducing NCoR levels after 6 h. These findings are
of importance in situations where enhancing progesterone activity is desirable, for example in the management of
endometrial cancer, the promotion of endometrial receptivity or the maintenance of myometrial quiescence during
pregnancy.
~2 Citings
115. Glutamate receptors and the regulation of steroidogenesis in the human adrenal gland: The metabotropic
pathway
By Felizola, Saulo J. A.; Nakamura, Yasuhiro; Satoh, Fumitoshi; Morimoto, Ryo; Kikuchi, Kumi; Nakamura, Tomohiro;
Hozawa, Atsushi; Wang, Lin; Onodera, Yoshiaki; Ise, Kazue; et al
From Molecular and Cellular Endocrinology (2014), 382(1), 170-177. Language: English, Database: CAPLUS,
DOI:10.1016/j.mce.2013.09.025
L-Glutamate is a major excitatory neurotransmitter in the mammalian brain. Glutamate receptors have been reported in
the rat adrenal cortex and in human aldosterone-producing adenomas (APA). However, details regarding the expression
levels and functions of these receptors in human adrenocortical tissues remain unknown. The mRNA levels of glutamate
receptors were evaluated by qPCR in: 12 normal adrenal cortex (NAC), 11 APA, and 12 cortisol-producing adenoma
(CPA) tissues. Protein localization was evaluated by immunohistochem. for 15 NAC, 5 idiopathic hyperaldosteronism
cases, 15 APA and 15 CPA. H295R cells were treated with angiotensin-II or forskolin alone or combined with the
GRM2/3 agonist LY 354740. The level of GRM3 mRNA was higher in APA than in CPA or NAC. GRM1, IGLUR2, and
IGLUR3 were also detected in adrenocortical tissues. When added to angiotensin-II/forskolin treatments, LY 354740
decreased aldosterone and cortisol prodn. in H295R cells. GRM3 is considered to regulate steroidogenesis in
adrenocortical tissues.
~11 Citings
116. Hedgehog-signaling is upregulated in non-producing human adrenal adenomas and antagonism of

hedgehog-signaling inhibits proliferation of NCI-H295R cells and an immortalized primary human adrenal cell
line
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By Werminghaus, Pascal; Haase, Matthias; Hornsby, Peter J.; Schinner, Sven; Schott, Matthias; Malendowicz, Ludwik
K.; Lammers, Bernhard J.; Goretzki, Peter E.; Mueller-Mattheis, Volker; Markus Giessing; et al
Hedgehog (Hh)-signaling pathway is important in embryonic development. Activation of Hh-signaling is assocd. with
tumorigenesis. Recent studies demonstrate that Hh-signaling is involved in the development of the adrenal gland in mice
and is important in regulating adrenal proliferation. We studied the expression of Sonic hedgehog (SHH), Smoothened
(SMO), Patched1 (PTCH1) and GLI family zinc finger 1 (GLI1) in human adrenal and in adrenocortical tumors using
immunohistochem. and semi-quant. reverse transcriptase-polymerase chain reaction. Modulation of GLI1 and SMO
mRNA (mRNA) expression was investigated with forskolin. The role of Hh-signaling was studied in NCI-H295R cells and
in an immortalized primary cell line using the Hh-agonist smoothened agonist (SAG) and the Hh-antagonist cyclopamine.
The Hh-pathway components SHH, GLI1, PTCH1 and SMO were detectable in all adrenal glands. While in cortisol-
producing adenomas (CPA), Hh-signaling expression levels were comparable to that in normal adrenal cortex, a much
higher mRNA expression of GLI1, SMO and SHH was obsd. in non-producing adenomas (NPA). Interestingly,
stimulation of cultured adrenal cells with forskolin led to a decrease in expression of GLI1 and SMO mRNAs.
Antagonism of Hh-signaling resulted in a lower proliferation rate of adrenocortical cells, while Hh-agonism had no
significant effect on adrenal cell proliferation. Our data show Hh-signaling activity in adult adrenal glands. Activation of
the PKA pathway results in lower expression of Hh-signaling proteins. This might explain the lower expression of the Hh
components GLI1 and SMO in CPA in comparison to the higher expression in NPA. Hh-signaling might be involved in
the tumorigenesis of NPA.
~5 Citings
117. A Drug Repositioning Approach Identifies Tricyclic Antidepressants as Inhibitors of Small Cell Lung Cancer
and Other Neuroendocrine Tumors
By Jahchan, Nadine S.; Dudley, Joel T.; Mazur, Pawel K.; Flores, Natasha; Yang, Dian; Palmerton, Alec; Zmoos,
Anne-Flore; Vaka, Dedeepya; Tran, Kim Q. T.; Zhou, Margaret; et al
From Cancer Discovery (2013), 3(12), 1364-1377. Language: English, Database: CAPLUS, DOI:10.1158/2159-
8290.CD-13-0183
Small cell lung cancer (SCLC) is an aggressive neuroendocrine subtype of lung cancer with high mortality. We used a
systematic drug repositioning bioinformatics approach querying a large compendium of gene expression profiles to
identify candidate U.S. Food and Drug Administration (FDA)-approved drugs to treat SCLC. We found that tricyclic
antidepressants and related mols. potently induce apoptosis in both chemonaive and chemoresistant SCLC cells in
culture, in mouse and human SCLC tumors transplanted into immunocompromised mice, and in endogenous tumors
from a mouse model for human SCLC. The candidate drugs activate stress pathways and induce cell death in SCLC
cells, at least in part by disrupting autocrine survival signals involving neurotransmitters and their G protein-coupled
receptors. The candidate drugs inhibit the growth of other neuroendocrine tumors, including pancreatic neuroendocrine
tumors and Merkel cell carcinoma. These expts. identify novel targeted strategies that can be rapidly evaluated in
patients with neuroendocrine tumors through the repurposing of approved drugs.
~52 Citings
118. Compounds and methods of use thereof for treating neurodegenerative disorders
By Zack, Donald J.; Welsbie, Derek Stuart; Yang, Zhiyong
In some embodiments, the presently disclosed subject matter provides a method for treating or preventing a
neurodegenerative disease, disorder, or condition in a subject in need thereof, the method comprising administering to
the subject a 15 therapeutically effective amt. of a compd. of Formula I (wherein D is selected from the group consisting
of O, S(O)0-2, and NR1b; R1a is selected from the group consisting of H, substituted or unsubstituted alkyl, halogen, OR1b,
NO2, NR1bR1c; wherein: R1b and R1c are each independently selected from the group consisting of H or substituted or
unsubstituted alkyl, aryl, etc.; R1 is H, substituted alkyl, aryl, etc.; Q is CR2a or N wherein R2a is H, alkyl or cyano; Z is O,
NR1b and S(O)0-2; and Ar is substituted aryl or substituted heteroaryl), or a pharmaceutically acceptable salt, hydrate, or
prodrug thereof, thereby treating or preventing the neurodegenerative disease, disorder, or condition.
SciFinder® Page 48
~2 Citings
119. Crystal structure of a glucose/H+ symporter and its mechanism of action

By Iancu, Cristina V.; Zamoon, Jamillah; Woo, Sang Bum; Aleshin, Alexander; Choe, Jun-yong
From Proceedings of the National Academy of Sciences of the United States of America (2013), 110(44), 17862-
17867,S17862/1-S17862/15. Language: English, Database: CAPLUS, DOI:10.1073/pnas.1311485110
Glucose transporters are required to bring glucose into cells, where it is an essential energy source and precursor in
protein and lipid synthesis. These transporters are involved in important common diseases such as cancer and diabetes.
Here, we report the crystal structure of the Staphylococcus epidermidis glucose/H+ symporter in an inward-facing
conformation at 3.2-Å resoln. The Staphylococcus epidermidis glucose/H+ symporter is homologous to human glucose
transporters, is very specific and has high avidity for glucose, and is inhibited by the human glucose transport inhibitors
cytochalasin B, phloretin, and forskolin. On the basis of the crystal structure in conjunction with mutagenesis and
functional studies, we propose a mechanism for glucose/H+ symport and discuss the symport mechanism vs. facilitated
diffusion.
~30 Citings
120. Involvement of spinal PKA/CREB signaling pathway in the development of bone cancer pain
By Hang, Li-Hua; Yang, Jian-Ping; Shao, Dong-Hua; Chen, Zheng; Wang, Hong
From Pharmacological Reports (2013), 65(3), 710-716. Language: English, Database: CAPLUS, DOI:10.1016/S1734-
1140(13)71049-1
Background: It has been shown that spinal PKA/CREB signaling pathway is involved in neuropathic and inflammatory
pain, but its effects on bone cancer pain have not previously been investigated. The aim of this study was to examine
the potential role of the spinal PKA/CREB signaling pathway in the development of bone cancer pain. Methods: A bone
cancer pain model was made by inoculation of Walker 256 cells into the intramedullary space of rat tibia. Western blot
anal. examd. the expression of PKAca (PKA catalytic subunit) and phospho-CREB (p-CREB) protein levels. The authors
further investigated effects of intrathecal treatment with H-89 (a PKA inhibitor, 8 nmol) or forskolin (a PKA agonist, 10
nmol) on nociceptive behavior and the expression of PKAca and p-CREB. Results: On days 6, 9, and 15 after
inoculation, the expression of PKAca and p-CREB protein levels were higher in the bone cancer pain rats compared to
the sham rats. On day 9, intrathecal administration of H-89 significantly attenuated bone cancer-induced mech. allodynia
as well as upregulation of PKAca and p-CREB protein levels. These effects were completely abolished by intrathecal
pretreatment with the PKA agonist forskolin. Conclusion: The results suggest that the spinal PKA/CREB signaling
pathway may participate in the development of bone cancer pain. The findings of this study may provide an evidence for
developing novel analgesics to block bone cancer pain.
~9 Citings

SciFinder® Page 49
121. 11β-Hydroxyandrostenedione, the product of androstenedione metabolism in the adrenal, is metabolized in

LNCaP cells by 5α-reductase yielding 11β-hydroxy-5α-androstanedione
By Swart, Amanda C.; Schloms, Lindie; Storbeck, Karl-Heinz; Bloem, Liezl M.; du Toit, Therina; Quanson, Jonathan L.;
Rainey, William E.; Swart, Pieter
11β-Hydroxyandrostenedione (11OHA4), which is unique to the adrenal, was first isolated from human adrenal tissue in
the fifties. It was later shown in the sixties that 11β-hydroxytestosterone (11OHT) was also produced by the human
adrenal. Attention has shifted back to these adrenal androgens once more, as improved anal. techniques have enabled
more accurate detection of steroid hormones. In this paper, we investigated the origin of these metabolites as well as
their subsequent metab. and examd. a possible physiol. role for 11OHA4 in prostate cancer cells. In H295R cells treated
with forskolin and trilostane, etomidate, a reported cytochrome P 450 11β-hydroxylase (CYP11B1) inhibitor, blocked the
prodn. of corticosterone, cortisol, 11OHA4 and 11OHT. The metab. of androstenedione and testosterone by CYP11B1
and aldosterone synthase (CYP11B2) was assayed. Androstenedione was converted by CYP11B1, while the conversion
by CYP11B2 was negligible. Both enzymes readily converted testosterone. The metab. of these 11β-hydroxylated
metabolites by 11β-hydroxysteroid dehydrogenase (11βHSD) types 1 and 2 was subsequently investigated. 11βHSD2
catalyzed the conversion of both 11OHA4 and 11OHT to their resp. keto-steroids, while 11βHSD1 catalyzed the
conversion of 11-ketoandrostenedione and 11-ketotestosterone to their resp. hydroxy-steroids in Chinese hamster ovary
cells. Investigating a functional role, steroid 5α-reductase types 1 and 2 converted 11OHA4 to 11β-hydroxy-5α-
androstanedione (11OH-5α-dione), identified by accurate mass detection. UPLC-MS/MS analyses of 11OHA4 metab. in
LNCaP androgen-dependent prostate cancer cells, identified the 5α-reduced metabolite as well as 11-
ketoandrostenedione and 11-ketotestosterone, with the latter indicating conversion by 17β-hydroxysteroid
dehydrogenase. Downstream metab. by 11βHSD2 and by 5α-reductase may therefore indicate a physiol. role for
11OHA4 and/or 11OH-5α-dione in normal and prostate cancer cells.
~17 Citings
122. Preparation of small molecule modulators of the relaxin receptor 1 for treatment of cardiovascular and
other diseases
By Marugan, Juan Jose; Xiao, Jingbo; Ferrer-Alegre, Marc; Chen, Catherine; Southall, Noel; Zheng, Wei; Agoulnik,
Alexander; Agoulnik, Irina
Disclosed are small mol. modulators of the human relaxin
receptor 1 (RXFP1) useful in the treatment of RXFP1 mediated
facets of human health, e.g., cardiovascular disease. RXFP1
modulators of formula I [wherein A is (un)substituted 1,2-
phenylenyl, 1,2-heteroarylenyl, 1,2-heterocyclyl, etc.; R1 is -
NHCO(alkyl), Ph, O(alkyl), etc.; and R2 is (un)substituted alkyl
cycloalkyl, heteroarylalkyl, etc.] and their pharmaceutically
acceptable salts and compns. thereof are claimed and prepd.
For example, II was prepd. in a yield of 46% via the reaction of
2-(tert-butoxycarbonylamino)benzoic acid and 3-
(trifluoromethyl)aniline. I were assayed for RXFP1 modulating
activity measured by TR-FRET detection where II
demonstrated an AC50 of 5.29 µM and a maximal receptor
activation of 79% compared to forskolin.
~0 Citings

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123. CANCER-SPECIFIC GENE CARRIERS RESPONDING TO CANCER MICROENVIRONMENT: ACIDOSIS AND
HYPER-ACTIVATED PROTEIN KINASES
By Kushio, Satoshi; Tsuchiya, Akira; Nakamura, Yuta; Nobori, Takanobu; Kim, Chan Woo; Zhao, Guo Xi; Funamoto,
Taiki; Lee, Eun Kyung; Niidome, Takuro; Mori, Takeshi; et al
From Biomedical Engineering (Singapore, Singapore) (2013), 25(5), 1340005/1-1340005/11. Language: English,
Database: CAPLUS, DOI:10.4015/S101623721340005X
Protein kinase (PK)-responsive gene carriers modified with polyethylene glycol (PEG) chains using an acid-labile linker
were developed. These carriers were obtained by modifying the PEG chains and substrate peptides for the PKs (PKA or
PKCα) on the branched polyethyleneimine main chain. Polyplexes formed from these carriers and plasmid DNA (pDNA)
were stably dispersed under neutral pH medium. The polyplexes were also taken up by cells on the release of the PEG
chains under the slightly acidic extracellular pH assocd. with cancer cells. The polyplexes taken up by cells resulted in
gene expression when the substrate peptides were phosphorylated by the intracellular PKs to release pDNA from the
polyplexes. These novel gene carriers are expected to be promising for cancer-specific gene therapy via i.v.
administration.
~0 Citings

By Iancu, Cristina V.; Zamoon, Jamillah; Woo, Sang Bum; Aleshin, Alexander; Choe, Jun-yong
From Proceedings of the National Academy of Sciences of the United States of America, Early Edition (2013), (Oct 14
2013), 1-6, 6 pp.. Language: English, Database: CAPLUS, DOI:10.1073/pnas.1311485110
protein and lipid synthesis. These transporters are involved in important common diseases such as cancer and diabetes.
Here, we report the crystal structure of the Staphylococcus epidermidis glucose/H+ symporter in an inward-facing
conformation at 3.2-Å resoln. The Staphylococcus epidermidis glucose/H+ symporter is homologous to human glucose
transporters, is very specific and has high avidity for glucose, and is inhibited by the human glucose transport inhibitors
cytochalasin B, phloretin, and forskolin. On the basis of the crystal structure in conjunction with mutagenesis and
functional studies, we propose a mechanism for glucose/H+ symport and discuss the symport mechanism vs. facilitated
diffusion.
~0 Citings
125. Protein kinase C-induced activin A switches adrenocortical steroidogenesis to aldosterone by suppressing
CYP17A1 expression
By Hofland, Johannes; Steenbergen, Jacobie; Hofland, Leo J.; van Koetsveld, Peter M.; Eijken, Marco; van
Nederveen, Francien H.; Kazemier, Geert; de Herder, Wouter W.; Feelders, Richard A.; de Jong, Frank H.
From American Journal of Physiology (2013), 305(3, Pt. 1), E736-E744. Language: English, Database: CAPLUS,
DOI:10.1152/ajpendo.00034.2013
Functional zonation of the adrenal cortex is a consequence of the zone-specific expression of P450c17 (CYP17A1) and
its cofactors. Activin and inhibin peptides are differentially produced within the zones of the adrenal cortex and have
been implicated in steroidogenic control. In this study, we investigated whether activin and inhibin can function as
intermediates in functional zonation of the human adrenal cortex. Activin A suppressed CYP17A1 expression and
P450c17 function in adrenocortical cell lines as well as in primary adrenal cell cultures. Inhibin βA-subunit mRNA and
activin A protein levels were found to be increased up to 1900-fold and 49-fold, resp., after protein kinase C (PKC)
stimulation through PMA or angiotensin II in H295R adrenocortical carcinoma cells. This was confirmed in HAC15 cells
and for PMA in primary adrenal cell cultures. Both PMA and Ang II decreased CYP17A1 expression in the adrenocortical
cell lines, whereas PMA concurrently suppressed CYP17A1 levels in the primary cultures. Inhibition of activin signaling
during PKC stimulation through silencing of the inhibin βA-subunit or blocking of the activin type I receptor opposed the
PMA-induced downregulation of CYP17A1 expression and P450c17 function. In contrast, PKA stimulation through
ACTH or forskolin increased expression of the inhibin α-subunit and betaglycan, both of which are antagonists of activin
action. These data indicate that activin A acts as a PKC-induced paracrine factor involved in the suppression of
CYP17A1 in the zona glomerulosa and can thereby contribute to functional adrenocortical zonation.
~3 Citings
126. Cyclic AMP regulates the migration and invasion potential of human pancreatic cancer cells
SciFinder® Page 51
By Zimmerman, Noah P.; Roy, Ishan; Hauser, Andrew D.; Wilson, Jessica M.; Williams, Carol L.; Dwinell, Michael B.
From Molecular Carcinogenesis (2015), 54(3), 203-215. Language: English, Database: CAPLUS,
DOI:10.1002/mc.22091
Aggressive dissemination and metastasis of pancreatic ductal adenocarcinoma (PDAC) results in poor prognosis and
marked lethality. Rho monomeric G protein levels are increased in pancreatic cancer tissue. As the mechanisms
underlying PDAC malignancy are little understood, we investigated the role for cAMP in regulating monomeric G protein
regulated invasion and migration of pancreatic cancer cells. Treatment of PDAC cells with cAMP elevating agents that
activate adenylyl cyclases, forskolin, protein kinase A (PKA), 6-Bnz-cAMP, or the cyclic nucleotide phosphodiesterase
inhibitor cilostamide significantly decreased migration and Matrigel invasion of PDAC cell lines. Inhibition was dose-
dependent and not significantly different between forskolin or cilostamide treatment. CAMP elevating drugs not only
blocked basal migration, but similarly abrogated transforming-growth factor-β-directed PDAC cell migration and invasion.
The inhibitory effects of cAMP were prevented by the pharmacol. blockade of PKA. Drugs that increase cellular cAMP
levels decreased levels of active RhoA or RhoC, with a concomitant increase in phosphorylated RhoA. Diminished Rho
signaling was correlated with the appearance of thickened cortical actin bands along the perimeter of non-motile forskolin
or cilostamide-treated cells. Decreased migration did not reflect alterations in cell growth or programmed cell death.
Collectively these data support the notion that increased levels of cAMP specifically hinder PDAC cell motility through F-
actin remodeling.
~7 Citings
127. Aip regulates cAMP signalling and GH secretion in GH3 cells

By Formosa, R.; Xuereb-Anastasi, A.; Vassallo, J.
From Endocrine-Related Cancer (2013), 20(4), 495-505. Language: English, Database: CAPLUS, DOI:10.1530/ERC-
13-0043
Mutations in the aryl hydrocarbon receptor-interacting protein (AIP) gene have been linked to predisposition to pituitary
adenomas. However, the mechanism by which this occurs remains unknown. AIP interacts with a no. of interesting
proteins, including members of the cAMP signalling pathway that has been shown to be consistently altered in pituitary
tumors. The functional role of Aip was investigated using both over-expression and knock down of Aip in GH3 cells.
CAMP signalling and its downstream effectors, including GH secretion, were then investigated. CAMP signalling was
analyzed using cAMP assays, cAMP-response element-promoter luciferase reporter assays, real-time PCR and finally
secreted GH quantification. Over-expression of wild-type (WT)-Aip reduced forskolin-induced cAMP signalling at the
total cAMP level, luciferase reporter activity and target gene expression, when compared with empty vector and the non-
functional R304X mutant. Addnl., GH secretion was reduced in WT-Aip over-expressing GH3 cells treated with forskolin.
Knock down of endogenous Aip resulted in increased cAMP signalling but a decrease in GH secretion was also noted.
Inhibition of phosphodiesterase activity using general and selective inhibitors did not completely ablate the effect of Aip
on forskolin-augmented cAMP signalling. A mechanism by which Aip acts as a tumor suppressor, by maintaining a low
cAMP signalling and concn., is suggested. Mutations of Aip render the protein incapable of such activity. This effect
appears not to be mediated by the AIP-PDE interaction, suggesting the involvement of other interacting partners in
mediating this outcome.
~15 Citings
128. TSH induces metallothionein 1 in thyrocytes via Gq/11- and PKC-dependent signaling
By Baeck, Christer M.; Stohr, Stefanie; Schaefer, Eva A. M.; Biebermann, Heike; Boekhoff, Ingrid; Breit, Andreas;
Gudermann, Thomas; Buech, Thomas R. H.
From Journal of Molecular Endocrinology (2013), 51(1), 79-90. Language: English, Database: CAPLUS,
DOI:10.1530/JME-12-0200
Metallothioneins (MTs) are cytoprotective proteins acting as scavengers of toxic metal ions or reactive oxygen species.
MTs are upregulated in follicular thyroid carcinoma and are regarded as a marker of thyroid stress in Graves' disease.
However, the mechanism of MT regulation in thyrocytes is still elusive. In other cellular systems, cAMP-, calcium-, or
protein kinase C (PKC)-dependent signaling cascades have been shown to induce MT expression. Of note, all of these
three pathways are activated following the stimulation of the TSH receptor (TSHR). Thus, we hypothesized that TSH
represents a key regulator of MT expression in thyrocytes. In fact, TSHR stimulation induced expression of MT isoform
1X (MT1X) in human follicular carcinoma cells. In these cells, induction of MT1X expression critically relied on intact
Gq/11 signaling of the TSHR and was blocked by chelation of intracellular calcium and inhibition of PKC. TSHR-
independent stimulation of cAMP formation by treating cells with forskolin also led to an upregulation of MT1X, which was
completely dependent on PKA. However, inhibition of PKA did not affect the regulation of MT1X by TSH. As in follicular
thyroid carcinoma cells, TSH also induced MT1 protein in primary human thyrocytes, which was PKC dependent as well.
In summary, these findings indicate that TSH stimulation induces MT1X expression via Gq/11 and PKC, whereas cAMP-
PKA signaling does not play a predominant role. To date, little has been known regarding cAMP-independent effects of
TSHR signaling. Our findings extend the knowledge about the PKC-mediated functions of the TSHR.
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~1 Citing
129. Use of label-free waveguide grating biosensors to understand and identify treatment for cancer
By Deng, Huayun; Fang, Ye; Lahiri, Joydeep; Ling, Shizhang
From U.S. Pat. Appl. Publ. (2013), US 20130210057 A1 20130815, Language: English, Database: CAPLUS
The disclosure relates to methods of using dynamic mass redistribution data obtained from cancer cells cultured on
waveguide grating biosensors in the presence of agonists and in the presence of chemotherapeutic agents, for predicting
effective chemotherapies for the treatment of cancer.
~0 Citings
130. Rapgef2 connects GPCR-mediated cAMP signals to ERK activation in neuronal and endocrine cells
By Emery, Andrew C.; Eiden, Maribeth V.; Mustafa, Tomris; Eiden, Lee E.
From Science Signaling (2013), 6(281), 2003993/1-2003993/12, 13 pp.. Language: English, Database: CAPLUS,
DOI:10.1126/scisignal.2003993
G protein (heterotrimeric guanine nucleotide-binding protein)-coupled receptor (GPCR)-mediated increases in the second
messenger cyclic adenosine monophosphate (cAMP) activate the mitogen-activated protein kinase (MAPK) extracellular
signal-regulated kinase (ERK), and in neuroendocrine cells, this pathway leads to cAMP-dependent neuritogenesis
mediated through Rap1 and B-Raf. We found that the Rap guanine nucleotide exchange factor Rapgef2 was enriched
from primary bovine neuroendocrine cells by cAMP-agarose affinity chromatog. and that it was specifically eluted by
cAMP. With loss-of-function expts. in the rat neuronal cell line Neuroscreen-1 (NS-1) and gain-of-function expts. in
human embryonic kidney 293T cells, we demonstrated that Rapgef2 connected GPCR-dependent activation of adenylate
cyclase and increased cAMP concn. with the activation of ERK in neurons and endocrine cells. Furthermore, knockdown
of Rapgef2 blocked cAMP- and ERK-dependent neuritogenesis. Our data are consistent with a pathway involving the
cAMP-mediated activation of Rapgef2, which then stimulates Rap1, leading to increases in B-Raf, MEK, and ERK
activity.
~1 Citing
131. Discerning apical and basolateral properties of HT-29/B6 and IPEC-J2 cell layers by impedance
spectroscopy, mathematical modeling and machine learning
By Schmid, Thomas; Bogdan, Martin; Guenzel, Dorothee
From PLoS One (2013), 8(7), e62913. Language: English, Database: CAPLUS, DOI:10.1371/journal.pone.0062913
Quantifying changes in partial resistances of epithelial barriers in vitro is a challenging and time-consuming task in
physiol. and pathophysiol. Here, we demonstrate that elec. properties of epithelial barriers can be estd. reliably by
combining impedance spectroscopy measurements, math. modeling and machine learning algorithms. Conventional
impedance spectroscopy is often used to est. epithelial capacitance as well as epithelial and subepithelial resistance.
Based on this, the more refined two-path impedance spectroscopy makes it possible to further distinguish transcellular
and paracellular resistances. In a next step, transcellular properties may be further divided into their apical and
basolateral components. The accuracy of these derived values, however, strongly depends on the accuracy of the initial
ests. To obtain adequate accuracy in estg. subepithelial and epithelial resistance, artificial neural networks were trained
to est. these parameters from model impedance spectra. Spectra that reflect behavior of either HT-29/B6 or IPEC-J2
cells as well as the data scatter intrinsic to the used exptl. setup were created computationally. To prove the proposed
approach, reliability of the estns. was assessed with both modeled and measured impedance spectra. Transcellular and
paracellular resistances obtained by such neural network-enhanced two-path impedance spectroscopy are shown to be
sufficiently reliable to derive the underlying apical and basolateral resistances and capacitances. As an exemplary
perturbation of pathophysiol. importance, the effect of forskolin on the apical resistance of HT-29/B6 cells was quantified.
~1 Citing
132. Bronchorelaxation of the human bronchi by CFTR activators

SciFinder® Page 53
By Norez, Caroline; Jayle, Christophe; Becq, Frederic; Vandebrouck, Clarisse
From Pulmonary Pharmacology & Therapeutics (2014), 27(1), 38-43. Language: English, Database: CAPLUS,
DOI:10.1016/j.pupt.2013.06.008
The airway functions are profoundly affected in many diseases including asthma, COPD and cystic fibrosis (CF). CF the
most common lethal autosomal recessive genetic disease is caused by mutations of the CFTR (Cystic Fibrosis
transmembrane Conductance Regulator) gene, which normally encodes a multifunctional and integral membrane cAMP
regulated and ATP gated Cl- channel expressed in airway epithelial cells. Using human lung tissues obtained from
patients undergoing surgery for lung cancer, we demonstrated that CFTR participates in bronchorelaxation. Using
human bronchial smooth muscle cells (HBSMC), we applied iodide influx assay to analyze the CFTR-dependent ionic
transport and immunofluorescence technique to localize CFTR proteins. Moreover, the relaxation was studied in isolated
human bronchial segments after pre-contraction with carbachol to det. the implication of CFTR in bronchodilation.We
found in HBSMC that the pharmacol. and regulation of CFTR is similar to that of its epithelial counterpart both for
activation (using forskolin/genistein or a benzo[c]quinolizinium deriv.) and for inhibition (CFTRinh-172 and GPinh5a). With
human bronchial rings, we obsd. that whatever the compd. used including salbutamol, the activation of muscular CFTR
leads to a bronchodilation after constriction with carbachol. Altogether, these observations revealed that CFTR in the
human airways is expressed in bronchial smooth muscle cells and can be pharmacol. manipulated leading to the
hypothesis that this ionic channel could contribute to bronchodilation in human.
~4 Citings
133. CREB-Regulated Transcription Co-Activator Family Stimulates Promoter II-Driven Aromatase Expression in
Preadipocytes
By Samarajeewa, Nirukshi U.; Docanto, Maria M.; Simpson, Evan R.; Brown, Kristy A.
From Hormones & Cancer (2013), 4(4), 233-241. Language: English, Database: CAPLUS, DOI:10.1007/s12672-013-
0142-1
The dramatically increased prevalence of breast cancer after menopause is of great concern and is correlated with
elevated local levels of estrogens. This is mainly due to an increase in aromatase expression driven by its proximal
promoter II (PII). We have previously demonstrated that the CREB co-activator CRTC2 binds directly to PII and
stimulates its activity via mechanisms involving LKB1-AMPK in response to prostaglandin E2 (PGE2). There are three
members of the CRTC family (CRTC1-3) and this study aimed to characterize the role of other CRTCs in the activation of
aromatase PII. The expression and subcellular localization of CRTCs were examd. in preadipocytes using qPCR and
immunofluorescence. Under basal conditions, CRTC1 expression was the lowest, whereas CRTC3 transcripts were
present at higher levels. Basally, CRTC2 and CRTC3 were mainly cytoplasmic and PGE2 caused their nuclear
translocation. Reporter assays and chromatin immunopptn. (ChIP) were performed to assess the effect of CRTCs on PII
activity and binding. Basal PII activity was significantly increased with all CRTCs. Forskolin (FSK)/phorbol 12-myristate
13-acetate (PMA), to mimic PGE2, resulted in a further significant increase in PII activity with all CRTCs, with CRTC2 and
CRTC3 having greater effects. This was consistent with ChIP data showing an increased binding of CRTCs to PII with
FSK/PMA. Moreover, gene silencing of CRTC2 and CRTC3 significantly reduced the FSK/PMA-mediated stimulation of
aromatase activity. Interestingly, CRTCs acted cooperatively with CREB1 to increase PII activity, and both CREs were
found to be essential for the maximal induction of PII activity by CRTCs. Phosphorylation of CRTC2 at its AMPK target
site, Ser 171, dictated its subcellular localization, and the activation of aromatase PII in preadipocytes. In conclusion, this
study demonstrates that aromatase regulation in primary human breast preadipocytes involves more than one CRTC.
~10 Citings
134. Peptidic inhibitor of signal transmission from G-alpha-s (Gαs) to G-alpha-i (Gαi)-coupled receptor cascades
and their use in drug screening and therapeutic methods
By Stefan, Eduard; Bachmann, Verena; Huber, Roland
The invention relates to novel peptidic inhibitors of signal transmission from Gαs- to Gαi-coupled receptor cascades and
methods for their prophylactic use in cancer and hormone-related disease or heart failure.
~0 Citings
135. 1,3,5-Triazine-2-amine derivatives, their preparation, and their therapeutic and diagnostic application as
cannabinoid CB2 ligands
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By Arnaud, Joeelle; Artiaga, Martine; Barth, Francis; Hortala, Laurent; Martinez, Serge; Roux, Pascale
The invention is related to the prepn. of title compds. I [R1 = Ph
substituted one or more times with substituents independently
selected from a halogen, Alk, and OAlk; R2 = Ph substituted
one or more times with substituents independently selected
from NO2, CN, SO2NH2 and derivs., etc.; R3 = Alk; Alk = alkyl
optionally substituted with ≥1 halo atoms; and their acid addn.
salts] as agonists of CB2 cannabinoid receptors and their use
for treatment of the diseases it implies (no data). The invention
is also related to compds. I contg. one or more isotopes that
are compatible with their use as a marker in positron emission
tomog. or single-photon emission tomog. imaging, chosen from
carbon-11, fluorine-18 and iodine-123, for the diagnosis of
diseases assocd. with overexpression of the CB2 receptors (no
data). Thus, monoamination of 2,4-dichloro-6-methoxy-1,3,5-
triazine with 2-(4-fluorophenyl)ethanamine, Pd-coupling of 4-
chloro-N-[2-(4-fluorophenyl)ethyl]-6-methoxy-1,3,5-triazin-2-
amine with (3-methoxyphenyl)boronic acid and acidulation with
HCl (no data for the free base) gave II.bul.HCl (m.p. = 194°). I
have very good affinity in vitro for cannabinoid CB2 receptors
(IC50< 500 nM). The agonist nature of compds. I was
demonstrated by adenylate-cyclase inhibition models (no data).
The antagonist nature of compds. I was demonstrated in
models of reversion of the inhibition of adenylate cyclase
(stimulated by forskolin) induced by an agonist of CB2
receptors (no data). The inverse agonist nature of compds. I
was demonstrated in models of activation of adenylate cyclase
(stimulated by forskolin) (no data).
~0 Citings
136. 1,3,5-Triazine-2-amine derivatives, their preparation, and their therapeutic and diagnostic application as
SciFinder® Page 55
By Arnaud, Joelle; Artiaga, Martine; Barth, Francis; Hortala, Laurent; Martinez, Serge; Roux, Pascale
From Fr. Demande (2013), FR 2983859 A1 20130614, Language: French, Database: CAPLUS
The invention is related to the prepn. of title compds. I [R1 = Ph substituted one or more times with substituents
independently selected from a halogen, Alk, and OAlk; R2 = Ph substituted one or more times with substituents
independently selected from NO2, CN, SO2NH2 and derivs., etc.; R3 = Alk; Alk = alkyl optionally substituted with ≥1 halo
atoms; and their acid addn. salts] as agonists of CB2 cannabinoid receptors and their use for treatment of the diseases it
implies (no data). The invention is also related to compds. I contg. one or more isotopes that are compatible with their
use as a marker in positron emission tomog. or single-photon emission tomog. imaging, chosen from carbon-11, fluorine-
18 and iodine-123, for the diagnosis of diseases assocd. with overexpression of the CB2 receptors (no data). Thus,
monoamination of 2,4-dichloro-6-methoxy-1,3,5-triazine with 2-(4-fluorophenyl)ethanamine, Pd-coupling of 4-chloro-N-[2-
(4-fluorophenyl)ethyl]-6-methoxy-1,3,5-triazin-2-amine with (3-methoxyphenyl)boronic acid and acidulation with HCl (no
data for the free base) gave II.bul.HCl (m.p. = 194°). I have very good affinity in vitro for cannabinoid CB2 receptors
(IC50< 500 nM). The agonist nature of compds. I was demonstrated by adenylate-cyclase inhibition models (no data).
The antagonist nature of compds. I was demonstrated in models of reversion of the inhibition of adenylate cyclase
(stimulated by forskolin) induced by an agonist of CB2 receptors (no data). The inverse agonist nature of compds. I was
demonstrated in models of activation of adenylate cyclase (stimulated by forskolin) (no data).
~0 Citings
137. Phosphodiesterases-11 (PDE11) inhibitors for increasing cortisol prodn. and treating cancer and other
diseases such as autoimmune disorders
By Hoffman, Charles Stuart; Ceyhan, Ozge
The intervention generally relates to compns. comprising a PDE11 inhibitor for increasing cortisol levels in a subject, or
for increasing the prodn. of cortisol from adenocortical cells in a subject. Aspects of the invention relate to the use of a
PDE11 inhibitor in a method of treatment of low cortisol levels and/or adrenal insufficiency in a subject, or a method of
treating a disease or disorders assocd. with adrenal insufficiency. Aspects of the invention relate to PDE11 inhibitors
belonging to compds. of formula e.g. I [R1, R2, R3 = independently H, halo, aliph., hetero-aliph., acyl, aryl, heteroaryl etc.;
X = O, S, NR4; R4 = H, alkyl; m = 0-2], and their use alone or in combination with long-term corticosteroid treatment to
elevate cortisol levels in a subject, and/or to treat adrenal insufficiency. Another aspect relates to kits comprising PDE11
inhibitors for administration to a subject, to increase cortisol levels and/or treat adrenal insufficiency, and their use as an
adjuvant for treatment of cancer or other disorders such as psychiatric diseases where elevating cortisol levels would be
beneficial to the subject.
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~2 Citings
138. Ursodeoxycholic acid attenuates colonic epithelial secretory function

By Kelly, Orlaith B.; Mroz, Magdalena S.; Ward, Joseph B. J.; Colliva, Carolina; Scharl, Michael; Pellicciari, Roberto;
Gilmer, John F.; Fallon, Padraic G.; Hofmann, Alan F.; Roda, Aldo; et al
From Journal of Physiology (Oxford, United Kingdom) (2013), 591(9), 2307-2318. Language: English, Database:
CAPLUS, DOI:10.1113/jphysiol.2013.252544
Dihydroxy bile acids, such as chenodeoxycholic acid (CDCA), are well known to promote colonic fluid and electrolyte
secretion, thereby causing diarrhoea assocd. with bile acid malabsorption. However, CDCA is rapidly metabolised by
colonic bacteria to ursodeoxycholic acid (UDCA), the effects of which on epithelial transport are poorly characterised.
Here, we investigated the role of UDCA in the regulation of colonic epithelial secretion. Cl- secretion was measured
across voltage-clamped monolayers of T84 cells and muscle-stripped sections of mouse or human colon. Cell surface
biotinylation was used to assess abundance/surface expression of transport proteins. Acute (15 min) treatment of T84
cells with bilateral UDCA attenuated Cl- secretory responses to the Ca2+ and cAMP-dependent secretagogues carbachol
(CCh) and forskolin (FSK) to 14.0 ± 3.8 and 40.2 ± 7.4% of controls, resp. (n = 18, P < 0.001). Investigation of the mol.
targets involved revealed that UDCA acts by inhibiting Na+/K+-ATPase activity and basolateral K+ channel currents,
without altering their cell surface expression. In contrast, i.p. administration of UDCA (25 mg kg-1) to mice enhanced
agonist-induced colonic secretory responses, an effect we hypothesised to be due to bacterial metab. of UDCA to
lithocholic acid (LCA). Accordingly, LCA (50-200 µM) enhanced agonist-induced secretory responses in vitro and a
metabolically stable UDCA analog, 6α-methyl-UDCA, exerted anti-secretory actions in vitro and in vivo. In conclusion,
UDCA exerts direct anti-secretory actions on colonic epithelial cells and metabolically stable derivs. of the bile acid may
offer a new approach for treating intestinal diseases assocd. with diarrhoea.
~5 Citings
139. Increased Th17 Cells in the Tumor Microenvironment Is Mediated by IL-23 via Tumor-Secreted
Prostaglandin E2
By Qian, Xuesong; Gu, Ling; Ning, Huan; Zhang, Yanping; Hsueh, Eddy C.; Fu, Mingui; Hu, Xiaoyu; Wei, Lin; Hoft,
Daniel F.; Liu, Jianguo
From Journal of Immunology (2013), 190(11), 5894-5902. Language: English, Database: CAPLUS,
DOI:10.4049/jimmunol.1203141
Tumor cell-derived mols. such as cytokines and lipid mediators play a crit. role in inducing chronic inflammation in the
tumor microenvironment. We found that Th17 cells were increased in the peripheral blood, spleen, and tumor tissues of
mammary gland tumor-bearing mice. The Th17 cell survival factor, IL-23, was also overexpressed in tumor tissues
isolated from mice and human breast cancer patients. Sol. mols. secreted from breast tumor cells, but not normal breast
epithelial cells, induced IL-23 protein secretion in dendritic cells via induction of p19 mRNA expression. Our data further
indicate that tumor-secreted PGE2 through EP2 and EP4 receptors enhanced IL-23 p19 gene transcription through
binding to the cAMP-response element in the p19 promoter. Blocking PGE2 synthesis by NS398, a COX2 inhibitor,
abrogated the enhancement of p19 expression both in vitro and in vivo. Furthermore, blocking protein kinase A (PKA) by
H89 completely abrogated the inductive effects of tumor-conditioned medium and PGE2 on p19 transcription, whereas
the cAMP active analog, Forskolin, mimics the PGE2 effect. Taken together, our results indicate that tumor-secreted
PGE2 induces IL-23, but not IL-12, prodn. in the tumor microenvironment, leading to Th17 cell expansion. This inductive
effect of PGE2 on IL-23 p19 transcription is mediated through cAMP/PKA signaling transduction pathway.
~15 Citings

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140. Antagonists targeting abnormal vasopressin V2 receptor
By North, William G.; Pang, Roy H. L.
The authors disclose the use of antagonists of an abnormal vasopressin receptor V2 (e.g., AbnV2) for use in cancer cell
imaging, or diagnosis of cancers, and cancer therapy. In one example, an antagonistic anti-AbnV2 antibody is shown to
reduce cell viability and proliferation of small cell lung cancer cells.
~0 Citings
141. Provasopressin antagonists: cancer therapy and diagnosis

By North, William G.; Pang, Roy H. L.
The authors disclose antibodies and antigen-binding fragments specific for provasopressin (pro-VP) for use in identifying
and targeting pro-VP-expressing cancer cells. In one example, an anti-vasopressin-assocd. glycopeptide monoclonal
antibody is shown to inhibit breast cancer and ovarian cancer growth in xenograft models.
~0 Citings
142. Triple combination antitumor regimen for the treatment of cancer

By Heeschen, Christopher; Hermann, Patrick C.; Lonardo, Enza
The invention relates to a triple combination comprising a nodal/activin pathway inhibitor, a hedgehog pathway inhibitor
and a chemotherapeutic agent and its use as a medicament for the treatment of cancer. In another aspect of the
invention the nodal/ activin pathway inhibitor is selected from e.g. SB431542, cyclopamine, follistatin, arsenic trioxide
wherein the hedgehog pathway inhibitor is CUR199691. Chemotherapeutic agent is selected from the list comprising:
gemcitabine, capecitabine, methotrexate, oxaliplatin, cisplatin, carboplatin, cyclophosphamide, doxorubicin etc.
~1 Citing
143. Triple combination antitumor regimen for the treatment of cancer

By Heeschen, Christopher; Hermann, Patrick C.; Lonardo, Enza
From Eur. Pat. Appl. (2013), EP 2589385 A1 20130508, Language: English, Database: CAPLUS
The invention relates to a triple combination comprising a nodal/activin pathway inhibitor, a hedgehog pathway inhibitor
and a chemotherapeutic agent and its use as a medicament for the treatment of cancer. In another aspect of the
invention the nodal/ activin pathway inhibitor is selected from e.g. SB431542, cyclopamine, follistatin, arsenic trioxide
wherein the hedgehog pathway inhibitor is CUR199691. Chemotherapeutic agent is selected from the list comprising:
gemcitabine, capecitabine, methotrexate, oxaliplatin, cisplatin, carboplatin, cyclophosphamide, doxorubicin etc.
~3 Citings
144. Mitochondrial Complex I inhibitors and forced oxidative phosphorylation synergize in inducing cancer cell
death
By Palorini, Roberta; Simonetto, Tiziana; Cirulli, Claudia; Chiaradonna, Ferdinando
From International Journal of Cell Biology (2013), 243876, 14 pp.. Language: English, Database: CAPLUS,
DOI:10.1155/2013/243876
SciFinder® Page 58
Cancer cells generally rely mostly on glycolysis rather than oxidative phosphorylation (OXPHOS) for ATP prodn. In fact,
they are particularly sensitive to glycolysis inhibition and glucose depletion. On the other hand mitochondrial
dysfunctions, involved in the onset of the Warburg effect, are sometimes also assocd. with the resistance to apoptosis
that characterizes cancer cells. Therefore, combined treatments targeting both glycolysis and mitochondria function,
exploiting peculiar tumor features, might be lethal for cancer cells. In this study, we show that glucose deprivation and
mitochondrial Complex I inhibitors synergize in inducing cancer cell death. In particular, our results reveal that low doses
of Complex I inhibitors, ineffective on immortalized cells and in high glucose growth, become specifically cytotoxic on
cancer cells deprived of glucose. Importantly, the cytotoxic effect of the inhibitors on cancer cells is strongly enhanced
by forskolin, a PKA pathway activator, that we have previously shown to stimulate OXPHOS. Taken together, we
demonstrate that induction in cancer cells of a switch from a glycolytic to a more respirative metab., obtained by glucose
depletion or mitochondrial activity stimulation, strongly increases their sensitivity to low doses of mitochondrial Complex I
inhibitors. Our findings might be a valuable approach to eradicate cancer cells.
~4 Citings
145. Discovery, molecular and pharmacological characterization of GSA-10, a novel small-molecule positive
modulator of smoothened
By Gorojankina, Tatiana; Hoch, Lucile; Faure, Helene; Roudaut, Hermine; Traiffort, Elisabeth; Schoenfelder, Angele;
Girard, Nicolas; Mann, Andre; Manetti, Fabrizio; Solinas, Antonio; et al
DOI:10.1124/mol.112.084590
Activation of the Smoothened (Smo) receptor mediates Hedgehog (Hh) signaling. Hh inhibitors are in clin. trials for
cancer, and small-mol. Smo agonists may have therapeutic interests in regenerative medicine. Here, we have generated
and validated a pharmacophoric model for Smo agonists and used this model for the virtual screening of a library of com.
available compds. Among the 20 top-scoring ligands, we have identified and characterized a novel
quinolinecarboxamide deriv., Pr 4-(1-hexyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carboxamido) benzoate, (GSA-10),
as a Smo agonist. GSA-10 fits to the agonist pharmacophoric model with two hydrogen bond acceptor groups and four
hydrophobic regions. Using pharmacol., biochem., and mol. approaches, we provide compelling evidence that GSA-10
acts at Smo to promote the differentiation of multipotent mesenchymal progenitor cells into osteoblasts. However, this
mol. does not display the hallmarks of ref. Smo agonists. Remarkably, GSA-10 does not recognize the classic bodipy-
cyclopamine binding site. Its effect on cell differentiation is inhibited by Smo antagonists, such as MRT-83, SANT-1,
LDE225, and M25 in the nanomolar range, by GDC-0449 in the micromolar range, but not by cyclopamine and
CUR61414. Thus, GSA-10 allows the pharmacol. characterization of a novel Smo active site, which is notably not
targeted to the primary cilium and strongly potentiated by forskolin and cholera toxin. GSA-10 belongs to a new class of
Smo agonists and will be helpful for dissecting Hh mechanism of action, with important implications in physiol. and in
therapy.
~8 Citings
146. Transgenic Drosophila tumor stem cell model and method for selective inhibition of cancer stem cells in
refractory intestinal cancer or in relapse of intestinal cancer
By Perrimon, Norbert; Markstein, Michele
SciFinder® Page 59
The present invention provides a transgenic Drosophila tumor stem cell model and uses thereof. Also provided is a
method to selectively inhibit cancer stem cells in a mammal having refractory intestinal cancer or a relapse of intestinal
cancer. In one embodiment, the invention provides a transgenic Drosophila strain that allows for regulatable, e.g.,
inducible expression of an oncogene in stem cells, such as intestinal stem cells (ISCs), thereby providing a tumor stem
cell (TSC) model and an in vivo system for drug screening. In one embodiment, the present invention may provide a
system to study cellular and mol. aspects of ISCs under both normal homeostatic and aberrant oncogenic conditions
because Drosophila ISCs hold many parallels to their mammalian counterparts. In one embodiment, the invention
provides a transgenic Drosophila having a plurality of expression cassettes, e.g., stably integrated expression cassettes.
In one embodiment, a first expression cassette comprises a tissue- or cell lineage-specific transcription regulatory
element operably linked to an open reading frame encoding a gene product that regulates transcription of a specific
transcription regulatory element in trans, e.g., by binding that element (a driver). For instance, the driver may be a
transcription factor including one that is modified, e.g., truncated or substituted or fused to another mol., relative to a wild-
type transcription factor. In one embodiment, the driver does not regulate transcription regulatory elements in the
Drosophila genome that is the driver is a heterologous transcription factor or may be a modified Drosophila transcription
factor that does not bind endogenous (native) Drosophila genes. Another expression cassette in the plurality of
expression cassettes, e.g., a second expression cassette, comprises a transcription regulatory element that includes a
nucleotide sequence that binds the driver, which is operably linked to an open reading frame encoding an oncogene.
Oncogene expression is generally lethal and so the present system provides for upregulation of oncogene expression via
the binding of the driver to the nucleotide sequence that binds the driver, which is linked to the oncogene. A further
expression cassette, for instance, a third expression cassette, comprises a transcription regulatory element that includes
a nucleotide sequence that binds the driver, e.g., a nucleotide sequence that is identical to that in the second expression
cassette, which is operably linked to an open reading frame for a first marker gene, e.g., one that is optically detectable
(e.g., detectable by colorimetric, fluorescent or luminescent methods). Another one of the expression cassettes (e.g., a
fourth expression cassette) may comprise a transcription regulatory element that includes a sequence that binds the
driver, e.g., a nucleotide sequence that is identical to that in the second expression cassette or the third expression
cassette, or both, which is operably linked to an open reading frame for a second marker gene which is different than the
first marker gene. A fifth expression cassette may comprise a transcription regulatory element that is not regulated by
the driver but which is operably linked to an open reading frame encoding a gene product that regulates the driver or the
expression thereof, for instance, a conditional mutant of a competitor or repressor of the driver.
~0 Citings
147. Effect of polyphenols on production of steroid hormones from human adrenocortical NCI-H295R cells
By Hasegawa, Eri; Nakagawa, Saori; Sato, Momoe; Tachikawa, Eiichi; Yamato, Susumu
From Biological & Pharmaceutical Bulletin (2013), 36(2), 228-237. Language: English, Database: CAPLUS,
DOI:10.1248/bpb.b12-00627
Modulating steroid hormone levels is a curative and preventive measure for Cushing's syndrome, aldosteronism, and
various stress-triggered symptoms. Polyphenols have been reported to inhibit steroidogenic enzymes such as 3β-
hydroxysteroid dehydrogenase (3β-HSD) and aromatase. However, evidence for their inhibitory effects is fragmentary
because it has been detd. in studies with small groups of steroid hormones. To investigate the effects of steroids on
complete steroidogenic pathways, comprehensive anal. of steroid hormones is necessary. Here we cultured forskolin-
stimulated NCI-H295R, a human adrenocortical carcinoma cell line, in the presence of a polyphenol and employed GC-
MS to simultaneously det. the levels of nine steroid hormones (pregnenolone, progesterone, deoxycorticosterone,
aldosterone, 17α-hydroxyprogesterone, dehydroepiandrosterone, androstenedione, testosterone, and estradiol) in cell
culture supernatant. We found that daidzein, genistein, apigenin, hesperetin, naringenin, and eriodictyol significantly
reduced deoxycorticosterone and androstenedione levels, suggesting inhibition of 3β-HSD by these polyphenols.
Apigenin was more potent than other polyphenols in increasing the levels of pregnenolone and 17α-
hydroxyprogesterone, suggesting that it inhibits cytochrome P 450 (CYP) 17 and CYP21, as well as 3β-HSD. Real-time
reverse transcription polymerase chain reaction showed that apigenin significantly downregulated the expression levels
of 3β-HSD, CYP17, and CYP21 mRNA. This is the first study to demonstrate the inhibitory effects of apigenin on CYP17
and CYP21.
~7 Citings
148. Methadone but not Morphine Inhibits Lubiprostone-Stimulated Cl- Currents in T84 Intestinal Cells and
Recombinant Human ClC-2, but not CFTR Cl- Currents
By Cuppoletti, John; Chakrabarti, Jayati; Tewari, Kirti; Malinowska, Danuta H.
From Cell Biochemistry and Biophysics (2013), 66(1), 53-63. Language: English, Database: CAPLUS,
DOI:10.1007/s12013-012-9406-6
SciFinder® Page 60
In clin. trials, methadone, but not morphine, appeared to prevent beneficial effects of lubiprostone, a ClC-2 Cl- channel
activator, on opioid-induced constipation. Effects of methadone and morphine on lubiprostone-stimulated Cl- currents
were measured by short circuit current (Isc) across T84 cells. Whole cell patch clamp of human ClC-2 (hClC-2) stably
expressed in HEK293 cells and in a high expression cell line (HEK293EBNA) as well as human CFTR (hCFTR) stably
expressed in HEK293 cells was used to study methadone and morphine effects on recombinant hClC-2 and hCFTR Cl-
currents. Methadone but not morphine inhibited lubiprostone-stimulated Isc in T84 cells with half-maximal inhibition at
100 nM. Naloxone did not affect lubiprostone stimulation or methadone inhibition of Isc. Lubiprostone-stimulated Cl-
currents in hClC-2/HEK293 cells, but not forskolin/IBMX-stimulated Cl- currents in hCFTR/HEK293 cells, were inhibited
by methadone, but not morphine. HEK293EBNA cells expressing hClC-2 showed time-dependent, voltage-activated,
CdCl2-inhibited Cl- currents in the absence (control) and the presence of lubiprostone. Methadone, but not morphine,
inhibited control and lubiprostone-stimulated hClC-2 Cl- currents with half-maximal inhibition at 100 and 200-230 nM,
resp. Forskolin/IBMX-stimulated hClC-2 Cl- currents were also inhibited by methadone. Myristoylated protein kinase
inhibitor (a specific PKA inhibitor) inhibited forskolin/IBMX- but not lubiprostone-stimulated hClC-2 Cl- currents.
Methadone caused greater inhibition of lubiprostone-stimulated currents added before patching (66.1 %) compared with
after patching (28.7 %). Methadone caused inhibition of lubiprostone-stimulated Cl- currents in T84 cells and control;
lubiprostone- and forskolin/IBMX-stimulated recombinant hClC-2 Cl- currents may be the basis for reduced efficacy of
lubiprostone in methadone-treated patients.
~10 Citings
149. MTA3 regulates CGB5 and Snail genes in trophoblast

By Chen, Ying; Miyazaki, Jun; Nishizawa, Haruki; Kurahashi, Hiroki; Leach, Richard; Wang, Kai
CAPLUS, DOI:10.1016/j.bbrc.2013.02.102
Secreted by the placental trophoblast, human chorionic gonadotropin (hCG) is an important hormone during pregnancy
and is required for the maintenance of pregnancy. Previous studies have shown that dys-regulation of hCG expression
is assocd. with preeclampsia. However, the exact relationship between altered hCG levels and development of
preeclampsia is unknown. Metastasis assocd. protein 3 (MTA3), a chromatin remodeling protein, is abundantly
expressed in the placental trophoblasts, but its function is unknown. In breast cancer, MTA3 has been shown to repress
the expression of Snail and cell migration. However, whether MTA3 acts similarly in the trophoblast has not been
investigated. In the present study, we examd. the role of MTA3 in regulating the hCG β-subunit gene (gene name:
CGB5) and Snail expression in the trophoblast cell line, BeWo, as well as its relevance to the high hCG expression levels
seen in preeclampsia. First, we investigated MTA3 expression in preeclamptic placenta as compared to normal control
placenta via gene expression microarray and qRT-PCR and found that MTA3 was significantly down-regulated, whereas
both CGB5 and Snail were up-regulated in preeclamptic placenta. Secondly, we knocked down MTA3 gene in
trophoblast cell line BeWo and found Snail and hCG were both up-regulated, suggesting that MTA3 represses Snail and
hCG gene expression in trophoblasts. Next, we cloned the CGB5 and Snail promoters into the pGL3-basic vector
individually and found that silencing of MTA3 by siRNA resulted in an increase of both CGB5 and Snail promoter
activities. To confirm that this MTA3 inhibition is a direct effect, we performed a chromatin immune-pptn. (ChIP) assay
and found that MTA3 occupied the proximal promoter regions of both Snail and hCG within BeWo cells. Furthermore, we
examd. MTA3 expression in placental trophoblast by immunohistochem. and found that MTA3 expression was higher in
villous cytotrophoblasts vs. syncytiotrophoblasts, which supports an inverse assocn. of MTA3 with hCG expression.
Lastly, using the well-characterized trophoblast fusion model, we examd. MTA3 and hCG levels in forskolin-treated
BeWo cells and found that MTA3 down-regulation was accompanied by an up-regulation of hCG. These data further
suggest that MTA3 is repressing placental hCG expression. In summary, MTA3 plays a crit. role in repressing hCG and
Snail in placenta trophoblast and its deregulation is assocd. with preeclampsia.
~5 Citings
150. Methods of treating and preventing mucoepidermoid carcinoma using imidazoquinolines

By Griffin, James D.; Wu, Lizi; Chen, Jie
SciFinder® Page 61
Imidazoquinolines, as set forth in formula I wherein R1 and R2
are each independently Me or ethyl; R3 is lower alkyl; and R4 is
pyridyl unsubstituted or substituted by halogen, cyano, lower
alkyl, lower alkoxy or piperazinyl unsubstituted or substituted by
lower alkyl; pyrimidinyl unsubstituted or substituted by lower
alkoxy; quinolinyl unsubstituted or substituted by halogen; or
quinoxalinyl; or a pharmaceutically acceptable salt thereof, are
useful for inhibiting growth or proliferation of mucoepidermoid
carcinoma cells.
~0 Citings
151. Selective inhibition of cell death in malignant vs normal B-cell precursors: implications for cAMP in
development and treatment of BCP-ALL
By Naderi, Elin Hallan; Ugland, Hege Katrin; Diep, Phoi-Phoi; Josefsen, Dag; Ruud, Ellen; Naderi, Soheil; Blomhoff,
Heidi Kiil
From Blood (2013), 121(10), 1805-1813. Language: English, Database: CAPLUS, DOI:10.1182/blood-2012-08-452698
B-cell precursor acute lymphoblastic leukemia (BCP-ALL) is the most commonly occurring pediatric cancer. Despite its
relatively good prognosis, there is a steady search for strategies to improve treatment effects and prevent the undesired
side effects on normal cells. In the present paper, we demonstrate a differential effect of cyclic adenosine
monophosphate (cAMP) signaling between normal BCPs and BCP-ALL blasts, pointing to a potential therapeutic window
allowing for manipulation of cAMP signaling in the treatment of BCP-ALL. By studying primary cells collected from
pediatric BCP-ALL patients and healthy controls, we found that cAMP profoundly decreased basal and DNA damage-
induced p53 levels and cell death in malignant cells, whereas normal BCP counterparts displayed slightly augmented cell
death when exposed to cAMP-increasing agents. We did not find evidence for a selection process involving generation
of increased basal cAMP levels in BCP-ALL cells, but we demonstrate that paracrine signaling involving prostaglandin
E2-induced cAMP generation has the potential to suppress p53 activation and cell death induction. The selective
inhibitory effect of cAMP signaling on DNA damage-induced cell death in BCP-ALL cells appears to be an acquired trait
assocd. with malignant transformation, potentially allowing the use of inhibitors of this pathway for directed killing of the
malignant blasts.
~4 Citings
152. Prostaglandin E2 promotes liver cancer cell growth by the upregulation of FUSE-binding protein 1
expression
SciFinder® Page 62
By Ma, Juan; Chen, Meng; Xia, Shu-Kai; Shu, Wei; Guo, Yan; Wang, Yao-Hui; Xu, Yan; Bai, Xiao-Ming; Zhang, Li;
Zhang, Hai; et al
From International Journal of Oncology (2013), 42(3), 1093-1104. Language: English, Database: CAPLUS,
DOI:10.3892/ijo.2013.1782
Liver cancer is a common human cancer with a high mortality rate and currently there is no effective chemoprevention or
systematic treatment. Recent evidence suggests that prostaglandin E2 (PGE2) plays an important role in the occurrence
and development of liver cancer. However, the mechanisms through which PGE2 promotes liver cancer cell growth are
not yet fully understood. It has been reported that the increased expression of FUSE-binding protein 1 (FBP1)
significantly induces the proliferation of liver cancer cells. In this study, we report that PGE2 promotes liver cancer cell
growth by the upregulation of FBP1 protein expression. Treatment with PGE2 and the E prostanoid 3 (EP3) receptor
agonist, sulprostone, resulted in the time-dependent increase in FBP1 protein expression; sulprostone increased the
viability of the liver cancer cells. The protein kinase A (PKA) inhibitor, H89, and the adenylate cyclase (AC) inhibitor,
SQ22536, inhibited the cell viability accelerated by sulprostone. By contrast, the Gi subunit inhibitor, pertussis toxin
(PTX), exhibited no significant effect. Treatment with PGE2 and sulprostone caused a decrease in JTV1 protein
expression, blocked the binding of JTV1 with FBP1, which served as a mechanism for FBP1 degrdn., leading to the
decreased ubiquitination of FBP1 and the increase in FBP1 protein expression. Furthermore, H89 and SQ22536
prevented the above effects of JTV1 and FBP1 induced by PGE2 and sulprostone. These findings indicate that the EP3
receptor activated by PGE2 may couple to Gs protein and activate cAMP (cAMP)-PKA, downregulating the levels of
JTV1 protein, consequently inhibiting the ubiquitination of FBP1 and increasing FBP1 protein expression, thus promoting
liver cancer cell growth. These observations provide new insights into the mechanisms through which PGE2 promotes
cancer cell growth.
~11 Citings
153. Protection against radiation-induced damage of 6-propyl-2-thiouracil (PTU) in thyroid cells

By Perona, Marina; Dagrosa, Maria A.; Pagotto, Romina; Casal, Mariana; Pignataro, Omar P.; Pisarev, Mario A.;
Juvenal, Guillermo J.
From Radiation Research (2013), 179(3), 352-360. Language: English, Database: CAPLUS, DOI:10.1667/RR2658.1
Many epidemiol. studies have shown that the exposure to high external radiation doses increases thyroid neoplastic
frequency, esp. when given during childhood or adolescence. The use of radioprotective drugs may decrease the
damage caused by radiation therapy and therefore could be useful to prevent the development of thyroid tumors. The
aim of this study was to investigate the possible application of 6-propyl-2-thiouracil (PTU) as a radioprotector in the
thyroid gland. Rat thyroid epithelial cells (FRTL-5) were exposed to different doses of γ irradn. with or without the addn.
of PTU, methimazole (MMI), reduced glutathione (GSH) and perchlorate (KClO4). Radiation response was analyzed by
clonogenic survival assay. CAMP (cAMP) levels were measured by RIA (RIA). Apoptosis was quantified by nuclear cell
morphol. and caspase 3 activity assays. Intracellular reactive oxygen species (ROS) levels were measured using the
fluorescent dye 2',7'-dichlorofluorescein-diacetate. Catalase, superoxide dismutase and glutathione peroxidase activities
were also detd. Pretreatment with PTU, MMI and GSH prior to irradn. significantly increased the surviving cell fraction
(SF) at 2 Gy (P < 0.05), while no effect was obsd. with KClO4. An increase in extracellular levels of cAMP was found
only in PTU treated cells in a dose and time-dependent manner. Cells incubated with agents that stimulate cAMP
(forskolin and dibutyril cAMP) mimicked the effect of PTU on SF. Moreover, pretreatment with the inhibitor of protein
kinase A, H-89, abolished the radioprotective effect of PTU. PTU treatment diminished radiation-induced apoptosis and
protected cells against radiation-induced ROS elevation and suppression of the antioxidant enzyme's activity. PTU was
found to radioprotect normal thyroid cells through cAMP elevation and redn. in both apoptosis and radiation-induced
oxidative stress damage.
~1 Citing
154. 5-Aza-2'-Deoxycytidine Has Minor Effects on Differentiation in Human Thyroid Cancer Cell Lines, But
Modulates Genes That Are Involved in Adaptation In Vitro
By Dom, Genevieve; Galdo, Vanessa Chico; Tarabichi, Maxime; Tomas, Gil; Hebrant, Aline; Andry, Guy; De Martelar,
Viviane; Libert, Frederick; Leteurtre, Emmanuelle; Dumont, Jacques E.; et al
From Thyroid (2013), 23(3), 317-328. Language: English, Database: CAPLUS, DOI:10.1089/thy.2012.0388
SciFinder® Page 63
Background: In thyroid cancer, the lack of response to specific treatment, for example, radioactive iodine, can be caused
by a loss of differentiation characteristics of tumor cells. It is hypothesized that this loss is due to epigenetic
modifications. Therefore, drugs releasing epigenetic repression have been proposed to reverse this silencing. Methods:
We investigated which genes were reinduced in dedifferentiated human thyroid cancer cell lines when treated with the
demethylating agent 5-aza-2'-deoxycytidine (5-AzadC) and the histone deacetylase inhibitors trichostatin A (TSA) and
suberoylanilide hydroxamic acid, by using reverse transcriptase-polymerase chain reaction and microarrays. These
results were compared to the expression patterns in in vitro human differentiated thyrocytes and in in vivo
dedifferentiated thyroid cancers. In addn., the effects of 5-AzadC on DNA quantities and cell viability were investigated.
Results: Among the canonical thyroid differentiation markers, most were not, or only to a minor extent, re-expressed by
5-AzadC, whether or not combined with TSA or forskolin, an inducer of differentiation in normal thyrocytes. Furthermore,
5-AzadC-modulated overall mRNA expression profiles showed only few commonly regulated genes compared to
differentiated cultured primary thyrocytes. In addn., most of the commonly strongly 5-AzadC-induced genes in cell lines
were either not regulated or upregulated in anaplastic thyroid carcinomas. Further anal. of which genes were induced by
5-AzadC showed that they were involved in pathways such as apoptosis, antigen presentation, defense response, and
cell migration. A no. of these genes had similar expression responses in 5-AzadC-treated nonthyroid cell lines.
Conclusions: Our results suggest that 5-AzadC is not a strong inducer of differentiation in thyroid cancer cell lines. Under
the studied conditions and with the model used, 5-AzadC treatment does not appear to be a potential redifferentiation
treatment for dedifferentiated thyroid cancer. However, this may reflect primarily the inadequacy of the model rather than
that of the treatment. Moreover, the observation that 5-AzadC neg. affected cell viability in cell lines could still suggest a
therapeutic opportunity. Some of the genes that were modulated by 5-AzadC were also induced in nonthyroid cancer
cell lines, which might be explained by an epigenetic modification resulting in the adaptation of the cell lines to their
culture conditions.
~3 Citings
155. Linking αMSH with PPARγ in B16-F10 melanoma

By Maresca, Vittoria; Flori, Enrica; Camera, Emanuela; Bellei, Barbara; Aspite, Nicaela; Ludovici, Matteo; Catricala,
Caterina; Cardinali, Giorgia; Picardo, Mauro
From Pigment Cell & Melanoma Research (2013), 26(1), 113-127. Language: English, Database: CAPLUS,
DOI:10.1111/j.1755-148X.2012.01042.x
We have discovered a new α-MSH (α-MSH)/peroxisome proliferator activated receptor-γ (PPAR-γ) connection in B16-
F10 cells. Both PPAR-γ up-regulation and its induction as an active transcription factor were obsd. in response to α-
MSH. The α-MSH/PPAR-γ connection influenced both pigmentation and proliferation. The forskolin-stimulated
cAMP/PKA pathway was not able to induce either PPAR-γ translocation into the nucleus or PPAR-γ transcriptional
activity. As the melanocortin-1 receptor, the specific receptor for the α-MSH, is a G-protein coupled receptor, we
wondered whether the phosphatidylinositol [PI(4,5)P2/PLCβ] signal pathway was involved in mediating the α-MSH-
dependent PPAR-γ activation. Employing inhibitors of PI(4,5)P2/PLCβ pathway, the results of our expts. suggested that
this pathway was promoted by α-MSH and that α-MSH played a role in mediating PPAR-γ activation. We have
demonstrated, for the first time, that α-MSH induces the PI(4,5)P2/PLCβ pathway, through anal. of the basic steps of the
pathway. The α-MSH effect on PPAR-γ was independent of animal species and was not correlated with the physio-
pathol. status.
~5 Citings
156. Protein kinase A (PKA) pathway is functionally linked to androgen receptor (AR) in the progression of
prostate cancer
By Sarwar, Martuza; Sandberg, Sabina; Abrahamsson, Per-Anders; Persson, Jenny L.
From Urologic Oncology: Seminars and Original Investigations (2014), 32(1), 25.e1-25.e12. Language: English,
Database: CAPLUS, DOI:10.1016/j.urolonc.2012.08.019
SciFinder® Page 64
Objectives: In the present study, we investigated whether the cyclic adenosine monophosphate (cAMP)-activated protein
kinase A (PKA) pathway may regulate the expression of AR and prostate-specific antigen (PSA) and whether there is a
correlation between the expression of cAMP/PKA-assocd. genes and androgen receptor (AR) in patients with prostate
cancer (CaP). Materials and methods: The functional studies were performed in LNCaP and PC3 cell lines. Data on the
mRNA expression of sets of genes in human clin. samples, including prostate tissues from organ donors, prostate
primary cancer, and metastatic cancer, were extd. from the National Center for Biotechnol. Informations Gene
Expression Omnibus (GEO) database. Statistical tests were applied. Results: We showed that elevated levels of
cAMP/PKA pathways induced an increased expression of AR and PSA proteins in LNCaP cells in the absence of
androgen. A cAMP-assocd. phosphodiesterase-4 (PDE4) inhibitor, rolipram induced an up-regulation in AR expression,
whereas a cAMP enhancer, forskolin increased PSA level without affecting AR expression. Forskolin treatment
increased the level of PKA R1α in LNCaP cells, but remarkably inhibited R1α expression in aggressive PC3 cells. In
patients with CaP, we found that the expression of genes encoding R1α and phosphodiesterase-4B was statistically
significantly lower in the metastatic specimens than that in the primary CaP specimens or in the normal prostate tissues
(P<0.01) and was reversely correlated with AR expression. Conversely, AR and PRKAR2B mRNA expressions were
significantly higher in metastatic lesions than those in the primary CaP specimens or in the normal prostate tissues
(P<0.01). Conclusion: Our study revealed a novel mechanism to precisely define the functional and clin. interrelationship
between the cAMP/PKA pathway and AR signaling in the development of androgen-independent growth of CaPs and
metastasis progression.
~5 Citings
157. Prostaglandin E2 induces stromal cell-derived factor-1 expression in prostate stromal cells by activating
protein kinase A and transcription factor Sp1
By Peng, Yanfei; Shi, Jiandang; Du, Xiaoling; Wang, Liang; Klocker, Helmut; Mo, Linjian; Mo, Zengnan; Zhang, Ju
From International Journal of Biochemistry & Cell Biology (2013), 45(3), 521-530. Language: English, Database:
CAPLUS, DOI:10.1016/j.biocel.2012.11.017
Recent reports indicate prostaglandin E2 (PGE2) can modulate tumor environment and promote angiogenesis through
induction of stromal cell-derived factor 1 (SDF-1) prodn. We investigated the mechanism of PGE2-induced SDF-1
regulation in human prostate stromal cell and analyzed the effects in a stromal-epithelial interaction model. PGE2
stimulation increased SDF-1 expression in the prostate stromal cell lines WPMY-1 and NAF. We revealed signaling
through the PGE2 receptor EP3 and activation of protein kinase A (PKA) are required. The EP3 agonist sulprostone and
the cAMP analog forskolin mimicked and the EP3 siRNA, antagonist L 798106 and the PKA inhibitor H 89 abrogated the
effect of PGE2 on SDF-1 expression. SDF-1 promoter truncation expts. demonstrated a 254 bp (from nt -219 to nt +34)
SDF-1 proximal promoter fragment contg. 5 putative transcription factor Sp1 binding motifs is sufficient for PGE2
induction. CHIP assays confirmed binding and PGE2 induced recruitment of Sp1 to the SDF-1 promoter. Sp1 motif
mutation identified Sp1 motifs -140/-133 and -9/+1 as the crucial elements responsible for PGE2 induction. Moreover,
SDF-1 was up- or down-regulated by Sp1 over-expression or knock-down. We also demonstrate stimulation of migration
of prostate cancer cell lines PC3 and DU145 with conditioned media collected from WPMY-1 or NAF cells stimulated with
PGE2 and blockade of enhanced migration by a SDF-1 neutralizing antibody. In conclusion, we provide evidence for a
paracrine prostate stromal-epithelial interaction induced by upregulation of expression of SDF-1 by PGE2. Our research
provides new insights into the mechanism promoting metastasis of prostate carcinoma via stromal-epithelial interaction.
~5 Citings
158. Canonical and noncanonical Hedgehog pathway in the pathogenesis of multiple myeloma
By Blotta, Simona; Jakubikova, Jana; Calimeri, Teresa; Roccaro, Aldo M.; Amodio, Nicola; Azab, Abdel Kareem;
Foresta, Umberto; Mitsiades, Constantine S.; Rossi, Marco; Todoerti, Katia; et al
From Blood (2012), 120(25), 5002-5013. Language: English, Database: CAPLUS, DOI:10.1182/blood-2011-07-368142
The Hedgehog (Hh) pathway is required for cell-fate detn. during the embryonic life, as well as cell growth and
differentiation in the adult organism, where the inappropriate activation has been implicated in several cancers. Here we
demonstrate that Hh signaling plays a significant role in growth and survival of multiple myeloma (MM) cells. We obsd.
that CD138+ MM cells express Hh genes and confirmed Smoothened (Smo)-dependent Hh signaling in MM using a
novel synthetic Smo inhibitor, NVP-LDE225 (Novartis), which decreased MM cell viability by inducing specific down-
regulation of Gli1 and Ptch1, hallmarks of Hh activity. In addn., we detected a nuclear localization of Gli1 in MM cells,
which is completely abrogated by Forskolin, a Gli1-modulating compd., confirming Smo-independent mechanisms
leading to Hh activation in MM. Finally, we identified that bone marrow stromal cells are a source of the Shh ligand,
although they are resistant to the Hh inhibitor because of defective Smo expression and Ptch1 up-regulation. Further in
vitro as well as in vivo studies showed antitumor efficacy of NVP-LDE225 in combination with bortezomib. Altogether,
our data demonstrate activation of both canonical and noncanonical Hh pathway in MM, thus providing the rationale for
testing Hh inhibitors in clin. trials to improve MM patient outcome.
SciFinder® Page 65
~25 Citings
159. FOXO1 controls thyroid cell proliferation in response to TSH and IGF-I and is involved in thyroid
tumorigenesis
By Zaballos, Miguel A.; Santisteban, Pilar
From Molecular Endocrinology (2013), 27(1), 50-62. Language: English, Database: CAPLUS, DOI:10.1210/me.2012-
1032
TSH and insulin/IGF-I synergistically induce the proliferation of thyroid cells mainly through the cAMP and
phosphatidylinositol 3-kinase (PI3K) pathways. However, the events involved in this cooperative induction remain
unknown, and mols. that are potentially controlled by both TSH and IGF-I are interesting candidates as integrators of
both stimuli. The finding that the PI3K pathway is frequently activated in thyroid malignancies has attracted attention to
this pathway in the thyroid field. One of the targets of PI3K is Forkhead box O (FoxO)-1, a widely expressed transcription
factor involved in a variety of cellular processes such as differentiation, proliferation, and apoptosis. Here we show that
FoxO1 is highly expressed in differentiated rat thyroid cells and human thyroid tissue compared with human thyroid
tumor-derived cells and surgically removed thyroid tumors, in which its expression is reduced. In differentiated cells,
TSH/cAMP treatment decreases FoxO1 mRNA and protein levels through proteasome activation, whereas both TSH and
IGF-I control FoxO1 localization by promoting a rapid exclusion from the nucleus in an Akt-dependent manner. FoxO1
can control p27KIP1 expression in differentiated and tumor cells of the thyroid. Furthermore, FoxO1 reexpression in tumor
cells promotes a decrease in their proliferation rate, whereas FoxO1 interference in differentiated cells increases their
proliferation. These data point to an important role of FoxO1 in mediating the effects of TSH and IGF-I on thyroid cell
proliferation and provide a link between loss of FoxO1 expression and the uncontrolled proliferation of thyroid tumor cells.
~8 Citings
160. Protein kinase A activation inhibits oncogenic Sonic hedgehog signalling and suppresses basal cell
carcinoma of the skin
By Makinodan, Eri; Marneros, Alexander G.
From Experimental Dermatology (2012), 21(11), 847-852. Language: English, Database: CAPLUS,
DOI:10.1111/exd.12016
Basal cell carcinoma of the skin (BCC) is caused by constitutive activation of the Sonic hedgehog (Shh) pathway, mainly
through mutations either in the Shh receptor Patched (PTCH) or in its co-receptor Smoothened (Smo). Inhibitors of this
pathway that are currently in clin. trials inhibit Smo. However, mutations in Smo can result in resistance to these
inhibitors. To target most BCCs and avoid acquired resistance because of Smo mutations, inhibiting the Shh-pathway
downstream of Smo is crit. Attractive downstream targets would be at the level of Gli proteins, the transcriptional
activators of this pathway in BCCs. Previously it has been shown that Gli1 and Gli2, when phosphorylated by protein
kinase A (PKA), are targeted for proteosomal degrdn. Here we show that PKA activation via the cAMP agonist forskolin
is sufficient to completely abolish oncogenic Smo activity in vitro. In an inducible BCC mouse model due to a Smo
mutation that confers resistance to current Smo inhibitors, topical forskolin treatment significantly reduced Gli1 mRNA
levels and resulted in strongly suppressed BCC tumor growth. Our data show that forskolin inhibits the growth of even
those BCCs that are resistant to Smo inhibitors and provide a proof-of-principle framework for the development of
topically applied human skin-permeable novel pharmacol. inhibitors of oncogenic Shh-signaling through PKA activation.
~8 Citings
161. A3 adenosine receptor mediates apoptosis in 5637 human bladder cancer cells by Gq protein/PKC-
dependent AIF upregulation
By Kanno, Takeshi; Gotoh, Akinobu; Fujita, Yumiko; Nakano, Takashi; Nishizaki, Tomoyuki
From Cellular Physiology and Biochemistry (2012), 30(5), 1159-1168. Language: English, Database: CAPLUS,
DOI:10.1159/000343306
SciFinder® Page 66
Background/Aims: A3 adenosine receptor mediates apoptosis in a variety of cancer cells via diverse signaling pathways.
The present study was conducted to assess A3 adenosine receptor-mediated apoptosis in human bladder cancer cell
lines and to understand the underlying mechanism. Methods: Human bladder cancer cell lines such as 253J, 5637, KK-
47, TCCSUP, T24, and UMUC-3 cells were cultured. The siRNA to silence the A3 adenosine receptor-targeted gene
was constructed and transfected into cells. MTT assay, TUNEL staining, Western blotting, and real-time RT-PCR were
carried out. Results: For all the investigated cell types adenosine induced apoptosis in a concn. (0.01-10 mM)- and
treatment time (24-48 h)-dependent manner. Adenosine-induced 5637 cell death was significantly inhibited by the A3
adenosine receptor inhibitor MRS1191 or knocking-down A3 adenosine receptor, and the A3 adenosine receptor agonist
2-Cl-IB-MECA mimicked the adenosine effect. The adenosine effect was prevented by GF109203X, an inhibitor of
protein kinase C (PKC), but it was not affected by forskolin, an activator of adenylate cyclase. Adenosine-induced 5637
cell death, alternatively, was not inhibited by the pan-caspase inhibitor Z-VAD. Adenosine upregulated expression of
apoptosis-inducing factor (AIF), that is suppressed by knocking-down A3 adenosine receptor, and accumulated AIF in the
nucleus. Conclusion: The results of the present study show that adenosine induces 5637 cell apoptosis by upregulating
AIF expression via an A3 adenosine receptor-mediated Gq protein/PKC pathway.
~7 Citings
162. Inhibition of CIP2A determines erlotinib-induced apoptosis in hepatocellular carcinoma

By Yu, Hui-Chuan; Chen, Hui-Ju; Chang, Ya-Ling; Liu, Chun-Yu; Shiau, Chung-Wai; Cheng, Ann-Lii; Chen, Kuen-Feng
From Biochemical Pharmacology (Amsterdam, Netherlands) (2013), 85(3), 356-366. Language: English, Database:
CAPLUS, DOI:10.1016/j.bcp.2012.11.009
Erlotinib is a small-mol. inhibitor of epidermal growth factor receptor (EGFR). Here, we identify that cancerous inhibitor of
protein phosphatase 2A (CIP2A) is a major determinant mediating erlotinib-induced apoptosis in hepatocellular
carcinoma (HCC). Erlotinib showed differential effects on apoptosis in 4 human HCC cell lines. Erlotinib induced
significant apoptosis in Hep3B and PLC5 cell lines; however, Huh-7 and HA59 T cell lines showed resistance to erlotinib-
induced apoptosis at all tested doses. Down-regulation of CIP2A, a cellular inhibitor of protein phosphatase 2A (PP2A),
mediated the apoptotic effect of erlotinib in HCC. Erlotinib inhibited CIP2A in a dose- and time-dependent manner in all
sensitive HCC cells whereas no alterations in CIP2A were found in resistant cells. Overexpression of CIP2A upregulated
phospho-Akt and protected Hep3B cells from erlotinib-induced apoptosis. In addn., silencing CIP2A by siRNA restored
the effects of erlotinib in Huh-7 cells. Moreover, adding okadaic acid, a PP2A inhibitor, abolished the effects of erlotinib
on apoptosis in Hep3B cells; and forskolin, a PP2A agonist enhanced the effect of erlotinib in resistant HA59 T cells.
Combining Akt inhibitor MK-2206 with erlotinib restored the sensitivity of HA59 T cells to erlotinib. Furthermore, in vivo
xenograft data showed that erlotinib inhibited the growth of PLC5 tumor but had no effect on Huh-7 tumor. Erlotinib
downregulated CIP2A and upregulated PP2A activity in PLC5 tumors, but not in Huh-7 tumors. In conclusion, inhibition
of CIP2A dets. the effects of erlotinib on apoptosis in HCC. CIP2A may be useful as a therapeutic biomarker for
predicting clin. response to erlotinib in HCC treatment.
~14 Citings
163. Biomarkers for epithelial cancer diagnosis and treatment

By Pitari, Giovanni M.; Zuzga, David S.
Disclosed herein are methods for diagnosing the state of an epithelial tumor or tissue in a subject. Also disclosed are
methods of diagnosing tumor initiation in an epithelial tissue of a subject. Also disclosed are methods of detg. prognosis
of a subject with an epithelial tumor. These methods include, in part, measuring the amt. of pSer239-VASP, pSerl57-
VASP, and/or total VASP in a subject or in a tissue or tumor of a subject and comparing those levels to an appropriate
ref. level. This abstr. is intended as a scanning tool for purposes of searching in the particular art and is not intended to
be limiting of the present invention.
~0 Citings
164. Identification of sumoylation sites in CCDC6, the first identified RET partner gene in papillary thyroid
carcinoma, uncovers a mode of regulating CCDC6 function on CREB1 transcriptional activity
By Luise, Chiara; Merolla, Francesco; Leone, Vincenza; Paladino, Simona; Sarnataro, Daniela; Fusco, Alfredo; Celetti,
Angela
SciFinder® Page 67
CCDC6 was originally identified in chimeric genes as caused by chromosomal translocation involving the RET
protooncogene in some thyroid tumors. Recognized as a 65 kDa pro-apoptotic phosphoprotein, CCDC6 has been
enrolled as an ATM substrate that contribute to protect genome integrity by modulating PP4c activity in response to
genotoxic stress. Recently, CCDC6 has been identified as a repressor of CREB1-dependent transcription. Sumoylation
has emerged as an important mechanism in transcriptional control. Here, we report the identification and
characterization of three sites of sumoylation in CCDC6 (K74, K266 and K424) which are highly conserved in
vertebrates. We demonstrate that the post-translational modifications by SUMO2 constrain most of the CCDC6 protein
in the cytosol and affect its functional interaction with CREB1 with a decrease of CCDC6 repressive function on CREB1
transcriptional activity. Indeed, the impairment of functional outcome of sumoylated CCDC6 is obtained knocking down
all three the sumoylation sites. Interestingly, in thyroid cells the SUMO2-mediated CCDC6 post-translational
modifications are induced by Forskolin, a cAMP analog. Signal transduction via the cAMP pathway is known to be
ubiquitous and represents a major line of communication between many organisms and their environment. We believe
that CCDC6 could be an important player in the dynamics of cAMP signaling by fine regulating CREB1 transcriptional
activity in normal and transformed thyroid cells.
~1 Citing
165. Pigment-independent cAMP-mediated epidermal thickening protects against cutaneous UV injury by

keratinocyte proliferation
By Scott, Timothy L.; Christian, Perry A.; Kesler, Melissa V.; Donohue, Kevin M.; Shelton, Brent; Wakamatsu,
Kazumasa; Ito, Shosuke; D'Orazio, John
From Experimental Dermatology (2012), 21(10), 771-777. Language: English, Database: CAPLUS,
DOI:10.1111/exd.12012
The epidermis increases pigmentation and epidermal thickness in response to UV exposure to protect against UV-
assocd. carcinogenesis; however, the contribution of epidermal thickness has been debated. In a humanized skin
mouse model that maintains interfollicular epidermal melanocytes, we found that forskolin, a small mol. that directly
activates adenylyl cyclase and promotes cAMP generation, up-regulated epidermal eumelanin accumulation in fair-
skinned melanocortin-1-receptor (Mc1r)-defective animals. Forskolin-induced pigmentation was assocd. with a
reproducible expansion of epidermal thickness irresp. of melanization or the presence of epidermal melanocytes.
Rather, forskolin-enhanced epidermal thickening was mediated through increased keratinocyte proliferation, indirectly
through secreted factor(s) from cutaneous fibroblasts. We identified keratinocyte growth factor (Kgf) as a forskolin-
induced fibroblast-derived cytokine that promoted keratinocyte proliferation, as forskolin induced Kgf expression both in
the skin and in primary fibroblasts. Lastly, we found that even in the absence of pigmentation, forskolin-induced
epidermal thickening significantly diminished the amt. of UV-A and UV-B that passed through whole skin and reduced the
amt. of UV-B-assocd. epidermal sunburn cells. These findings suggest the possibility of pharmacol.-induced epidermal
thickening as a novel UV-protective therapeutic intervention, particularly for individuals with defects in pigmentation and
adaptive melanization.
~9 Citings
166. Phosphodiesterase 11A (PDE11A) gene defects in patients with ACTH-independent macronodular adrenal
hyperplasia (AIMAH): functional variants may contribute to genetic susceptibility of bilateral adrenal tumors
By Vezzosi, Delphine; Libe, Rossella; Baudry, Camille; Rizk-Rabin, Marthe; Horvath, Anelia; Levy, Isaac; Rene-Corail,
Fernande; Ragazzon, Bruno; Stratakis, Constantine A.; Vandecasteele, Gregoire; et al
From Journal of Clinical Endocrinology and Metabolism (2012), 97(11), E2063-E2069. Language: English, Database:
CAPLUS, DOI:10.1210/jc.2012-2275
Context: Phosphodiesterases (PDEs) are key regulatory enzymes of intracellular cAMP levels. PDE11A function has
been linked to predisposition to adrenocortical tumors. Objective: The aim of the study was to study the PDE11A gene in
a large cohort of patients with ACTH-independent macronodular adrenal hyperplasia (AIMAH) and in control subjects.
Design: The PDE11A entire coding region was sequenced in 46 patients with AIMAH and 192 controls. Two variants
found in AIMAH patients were transiently expressed in HEK 293 and adrenocortical H295R cells for further functional
studies. Results: The frequency of all PDE11A variants was significantly higher among patients with AIMAH (28%)
compared to controls (7.2%) (P = 5 × 10-5). Transfection of the two PDE11A variants found in AIMAH patients only
(D609N or M878V) showed that cAMP levels were higher, after forskolin stimulation, in cells transfected with the
PDE11A mutants, compared to cells transfected with the wild-type PDE11A in HEK 293 cells (P < 0.05). Moreover,
transfection with mutants PDE11A increased transcriptional activity of a cAMP-response element reporter construct
compared to wild-type PDE11A in HEK 293 cells (P < 0.0004 for D609N and P < 0.003 for M878V) and in the
adrenocortical H295R cells (P < 0.05 for D609N and M878V). In addn., anal. of cAMP levels in intact living culture cells
by fluorescence resonance energy transfer probes showed increased cAMP in forskolin-treated cells transfected with
PDE11A variants compared with wild-type PDE11A (P < 0.05). Conclusion: We conclude that PDE11A genetic variants
may increase predisposition to AIMAH.
SciFinder® Page 68
~18 Citings
167. Functional status and relationships of melanocortin 1 receptor signaling to the cAMP and extracellular
signal-regulated protein kinases 1 and 2 pathways in human melanoma cells
By Herraiz, Cecilia; Journe, Fabrice; Ghanem, Ghanem; Jimenez-Cervantes, Celia; Garcia-Borron, Jose C.
Melanocortin 1 receptor (MC1R), a major determinant of skin phototype frequently mutated in melanoma, is a Gs protein-
coupled receptor that regulates pigment prodn. in melanocytes. MC1R stimulation activates cAMP synthesis and the
extracellular signal-regulated (ERK) ERK1 and ERK2. In human melanocytes, ERK activation by MC1R relies on cAMP-
independent transactivation of the c-KIT receptor. Thus MC1R functional coupling to the cAMP and ERK pathways may
involve different structural requirements giving raise to biased effects of skin cancer-assocd. mutations. We evaluated
the impact of MC1R mutations on ERK activation, cAMP prodn. and agonist binding. We found that MC1R mutations
impair cAMP prodn. much more often than ERK activation, suggesting less stringent requirements for functional coupling
to the ERK pathway. We examd. the crosstalk of the cAMP and ERK pathways in HBL human melanoma cells (wild-type
for MC1R, NRAS and BRAF). ERK activation by constitutively active upstream effectors or pharmacol. inhibition had
little effect on MC1R-stimulated cAMP synthesis. High cAMP levels were compatible with normal ERK activation but,
surprisingly, the adenylyl cyclase activator forskolin abolished ERK activation by MC1R, most likely by a cAMP-
independent mechanism. These results indicate little crosstalk of the cAMP and ERK pathways in HBL melanoma cells.
Finally, we studied cAMP accumulation in a panel of 22 human melanoma cell lines stimulated with MC1R agonists or
forskolin. cAMP synthesis was often inhibited, even in cells wild-type for MC1R and NRAS. Therefore, the cAMP
pathway is more frequently impaired in melanoma than could be predicted by the MC1R or NRAS genotype.
~12 Citings
168. The Essential Role of Giα2 in Prostate Cancer Cell Migration

By Zhong, Miao; Clarke, Shineka; Vo, BaoHan T.; Khan, Shafiq A.
From Molecular Cancer Research (2012), 10(10), 1380-1388. Language: English, Database: CAPLUS,
DOI:10.1158/1541-7786.MCR-12-0219
Cell- and receptor-specific regulation of cell migration by Gi/oα-proteins remains unknown in prostate cancer cells. In the
present study, oxytocin (OXT) receptor was detected at the protein level in total cell lysates from C81 (an androgen-
independent subline of LNCaP), DU145 and PC3 prostate cancer cells, but not in immortalized normal prostate luminal
epithelial cells (RWPE1), and OXT-induced migration of PC3 cells. This effect of OXT has been shown to be mediated
by Gi/oα-dependent signaling. Accordingly, OXT inhibited forskolin-induced luciferase activity in PC3 cells that were
transfected with a luciferase reporter for cAMP activity. Although mRNAs for all three Giα isoforms were present in PC3
cells, Giα2 was the most abundant isoform that was detected at the protein level. Pertussis toxin (PTx) inhibited the
OXT-induced migration of PC3 cells. Ectopic expression of the PTx-resistant Giα2-C352G, but not wild-type Giα2,
abolished this effect of PTx on OXT-induced cell migration. The Giα2-targeting siRNA was shown to specifically reduce
Giα2 mRNA and protein in prostate cancer cells. The Giα2-targeting siRNA eliminated OXT-induced migration of PC3
cells. These data suggest that Giα2 plays an important role in the effects of OXT on PC3 cell migration. The Giα2-
targeting siRNA also inhibited EGF-induced migration of PC3 and DU145 cells. Expression of the siRNA-resistant Giα2,
but not wild type Giα2, restored the effects of EGF in PC3 cells transfected with the Giα2-targeting siRNA. In conclusion,
Giα2 plays an essential role in OXT and EGF signaling to induce prostate cancer cell migration. Mol Cancer Res;
10(10); 1380-8. ©2012 AACR.
~2 Citings
169. Increasing the oxygen load by treatment with myo-inositol trispyrophosphate reduces growth of colon
cancer and modulates the intestine homeobox gene Cdx2
By Derbal-Wolfrom, L.; Pencreach, E.; Saandi, T.; Aprahamian, M.; Martin, E.; Greferath, R.; Tufa, E.; Choquet, P.;
Lehn, J.-M.; Nicolau, C.; et al
From Oncogene (2013), 32(36), 4313-4318. Language: English, Database: CAPLUS, DOI:10.1038/onc.2012.445
SciFinder® Page 69
Preventing tumor neovascularisation is one of the strategies recently developed to limit the dissemination of cancer cells
and apparition of metastases. Although these approaches could improve the existing treatments, a no. of unexpected
neg. effects have been reported, mainly linked to the hypoxic condition and the subsequent induction of the pro-
oncogenic hypoxia inducible factor(s) resulting from cancer cells' oxygen starvation. Here, we checked in vivo on colon
cancer cells an alternative approach. It is based on treatment with myo-inositol trispyrophosphate (ITPP), a mol. that
leads to increased oxygenation of tumors. We provide evidence that ITPP increases the survival of mice in a model of
carcinomatosis of human colon cancer cells implanted into the peritoneal cavity. ITPP also reduced the growth of s.c.
colon cancer cells xenografted in nu/nu mice. In the s.c. tumors, ITPP stimulated the expression of the homeobox gene
Cdx2 that is crucial for intestinal differentiation and that also has an anti-tumoral function. On this basis, human colon
cancer cells were cultured in vitro in hypoxic conditions. Hypoxia was shown to decrease the level of Cdx2 protein,
mRNA and the activity of the Cdx2 promoter. This decline was unrelated to the activation of HIF1α and HIF2α by
hypoxia. However, it resulted from the activation of a phosphatidylinositol 3-kinases-like mitogen-activated protein kinase
pathway, as assessed by the fact that LY294002 and U0126 restored high Cdx2 expression in hypoxia. Corroborating
these results, U0126 recapitulated the increase of Cdx2 triggered by ITPP in s.c. colon tumor xenografts. The present
study provides evidence that a chem. compd. that increases oxygen pressure can antagonize the hypoxic setting and
reduce the growth of human colon tumors implanted in nu/nu mice.
~3 Citings
170. The tumor suppressor RASSF10 is upregulated upon contact inhibition and frequently epigenetically
silenced in cancer
By Richter, A. M.; Walesch, S. K.; Wuerl, P.; Taubert, H.; Dammann, R. H.
From Oncogenesis (2012), 1(June), e18/1-e18/11. Language: English, Database: CAPLUS,
DOI:10.1038/oncsis.2012.18
The Ras assocn. domain family (RASSF) comprises a group of tumor suppressors that are frequently epigeneticaily
inactivated in various tumor entities and linked to apoptosis, cell cycle control and microtubule stability. In this work, we
concd. on the newly identified putative tumor suppressor RASSF10. Methylation anal. reveals RASSF10 promoter
hypermethylation in lung cancer, head and neck (HN) cancer, sarcoma and pancreatic cancer. An increase in RASSF10
methylation from normal tissues, primary tumors to cancer cell lines was obsd. Methylation was reversed by 5-aza-2'-
deoxycytidine treatment leading to reexpression of RASSF10. We further show that overexpression of RASSF10
suppresses colony formation in cancer cell lines. In addn., RASSF10 is upregulated by cell-cell contact and regulated on
promoter level as well as endogenously by forskolin, protein kinase A (PKA) and activator Protein 1 (AP-1), linking
RASSF10 to the cAMP signaling pathway. Knockdown of the AP-1 member JunD interfered with contact inhibition
induced RASSF10 expression. In summary, we found RASSF10 to be epigeneticaily inactivated by hypermethylation of
its CpG island promoter in lung, HN, sarcoma and pancreatic cancer. Furthermore, our novel findings suggest that tumor
suppressor RASSF10 is upregulated by PKA and JunD signaling upon contact inhibition and that RASSF10 suppresses
growth of cancer cells.
~2 Citings
171. cAMP-dependent protein kinase is essential for hypoxia-mediated epithelial-mesenchymal transition,

migration, and invasion in lung cancer cells
By Shaikh, Dooniya; Zhou, Qiyuan; Chen, Tianji; Ibe, Joyce Christina F.; Raj, J. Usha; Zhou, Guofei
From Cellular Signalling (2012), 24(12), 2396-2406. Language: English, Database: CAPLUS,
DOI:10.1016/j.cellsig.2012.08.007
Lung cancer is the leading cause of cancer-related death worldwide. Hypoxia is known to increase cancer cell migration
and invasion. We have previously reported that hypoxia induces epithelial-mesenchymal transition (EMT) in lung cancer
cells. However, it is unknown whether hypoxia promotes lung cancer cell migration and invasion via EMT and whether
cAMP (cAMP) dependent protein kinase (PKA) plays a role in this process. We found that hypoxia increased PKA
activity and induced mRNA and protein expression of PKA catalytic subunit α (PKACA), and regulatory subunits R1A and
R1B. Knockdown of HIF-1/2α prevented hypoxia-mediated induction of PKACA mRNA expression and PKA activity.
Inhibition of PKA activity with chem. inhibitors prevented EMT induced by hypoxia and tumor growth factor β1. However,
activation of PKA by forskolin and 8-Br-cAMP did not induce EMT. Furthermore, treatment with H89 and knockdown of
PKACA prevented hypoxia-mediated, EMT, cell migration, and invasion, whereas overexpression of mouse PKACA
rescued hypoxia-mediated migration and invasion in PKACA deficient cancer cells. Our results suggest that hypoxia
enhances PKA activity by upregulating PKA gene expression in a HIF dependent mechanism and that PKA plays a key
role in hypoxia-mediated EMT, migration, and invasion in lung cancer cells.
~15 Citings

SciFinder® Page 70
172. Methods of treating cancer using 3-(5-amino-2-methyl-4-oxo-4h-quinazolin-3-yl)-piperidine-2,6-dione
By Muller, George W.; Schafer, Peter H.; Man, Hon-Wah; Zhang, Ling-Hua; Gandhi, Anita; Chopra, Rajesh
Provided herein are methods of treating, preventing and/or
managing cancers, which comprise administering to a patient
3-(5-amino-2-methyl-4-oxo-4H-quinazolin-3-yl)-piperidine-2,6-
dione, or an enantiomer or a mixt. of enantiomers thereof, or a
pharmaceutically acceptable salt, solvate, hydrate, co-crystal,
clathrate, or polymorph thereof.
~5 Citings
173. Deficiency of proton-sensing ovarian cancer G protein-coupled receptor 1 attenuates glucose-stimulated

insulin secretion
By Nakakura, Takashi; Mogi, Chihiro; Tobo, Masayuki; Tomura, Hideaki; Sato, Koichi; Kobayashi, Masaki; Ohnishi,
Hiroshi; Tanaka, Shigeyasu; Wayama, Mitsutoshi; Sugiyama, Tetsuya; et al
From Endocrinology (2012), 153(9), 4171-4180. Language: English, Database: CAPLUS, DOI:10.1210/en.2012-1164
Ovarian cancer G protein-coupled receptor 1 (OGR1) has been shown as a receptor for protons. In the present study,
we aimed to know whether OGR1 plays a role in insulin secretion and, if so, the manner in which it does. To this end, we
created OGR1-deficient mice and examd. insulin secretion activity in vivo and in vitro. OGR1 deficiency reduced insulin
secretion induced by glucose administered i.p., although it was not assocd. with glucose intolerance in vivo. Increased
insulin sensitivity and reduced plasma glucagon level may explain, in part, the unusual normal glucose tolerance. In vitro
islet expts. revealed that glucose-stimulated insulin secretion was dependent on extracellular pH and sensitive to OGR1;
insulin secretion at pH 7.4 to 7.0, but not 8.0, was significantly suppressed by OGR1 deficiency and inhibition of Gq/11
proteins. Insulin secretion induced by KCl and tolbutamide was also significantly inhibited, whereas that induced by
several insulin secretagogues, including vasopressin, a glucagon-like peptide 1 receptor agonist, and forskolin, was not
suppressed by OGR1 deficiency. The inhibition of insulin secretion was assocd. with the redn. of glucose-induced
increase in intracellular Ca2+ concn. In conclusion, the OGR1/Gq/11 protein pathway is activated by extracellular protons
existing under the physiol. extracellular pH of 7.4 and further stimulated by acidification, resulting in the enhancement of
insulin secretion in response to high glucose concns. and KCl.
~18 Citings
174. Compositions for controlling neuronal outgrowth comprising IGFBPL1 or agonists

By Chen, Dong Feng; Guo, Chenying
Disclosed is a method of promoting neuronal growth by
administering insulin-like growth factor binding protein-like 1
(IGFBPL1), or an agent that increases or stabilizes IGFBPL1
activity to a subject in need thereof, e.g., a subject in need of
treating optic nerve degeneration. The present invention is
based in part on the discovery that IGFBPL1 is a key factor in
promoting axon outgrowth in retinal ganglion cells (RGCs)
through the regulation of IGF-1 signaling pathway. Specifically,
the compds. described herein treat diseases or injuries that can
cause optic nerve degeneration or axon degeneration in the
peripheral and central nervous system (CNS), such as
traumatic brain injury, spinal cord injury, multiple sclerosis,
Alzheimer's disease, and Parkinson's disease.
SciFinder® Page 71
~0 Citings
175. Compounds and methods of use thereof for treating neurodegenerative disorders
By Zack, Donald J.; Berlinicke, Cynthia; Hackler, Laszlo; Yang, Zhiyong; Steiner, Joseph P.; Vojkovsky, Tomas;
Ferraris, Dana; Slusher, Barbara S.; Inglese, James
Compds., compns., kits and methods for treating conditions
related to neurodegeneration or ocular disease, are disclosed.
~1 Citing
176. Carotenoid particles and uses thereof

By Petyaev, Ivan
This invention relates to the incorporation of bioactive cargo mols. into particles with carotenoids, such as lycopene. The
incorporation of a cargo mol. into a carotenoid particle may for example increase the bioavailability of the cargo mol. in
the bloodstream compared to other delivery systems. Carotenoid particles as described herein may be useful in the
formulation of therapeutic and nutritional compds. for oral administration to individuals. Thus, lycopene particles
incorporating whey proteins were produced by mixing whey protein and Lycored oleoresin, spray drying the mixt., and
mixing with soy lecithin in vegetable oil. Administration of the particles significantly improved symptoms in coronary heart
disease patients pos. for anti-Chlamydia IgG and hypercholesterolemia.
SciFinder® Page 72
~3 Citings
177. Hydroxylated estrogens (2-OH-E2 AND 4-OH-E2) do not activate cAMP/PKA and ERK1/2 pathways activation
in a breast cancer MCF-7 cell line
By Kwiecinska, P.; Ptak, A.; Wrobel, A.; Gregoraszczuk, E. L.
From Endocrine Regulations (2012), 46(1), 3-12. Language: English, Database: CAPLUS,
DOI:10.4149/endo_2012_01_3
Objective: The current study was undertaken to det. the involvement of cAMP/PKA and MAPK-mediated signalling
pathways in the regulation of cell proliferation by hydroxylated metabolites of 17β-estradiol (E2). Methods: MCF-7,
human breast cancer cells, were cultured in phenol red-free DMEM and treated with 1 nM 2-OH-E2 or 4-OH-E2. E2 was
used as a pos. control. Cell proliferation was measured using the BrdU incorporation assay. Cellular levels of cAMP and
PKA were detd. using ELISA kits. ERK1/2 protein expression was evaluated by Western Blot anal. To det. the
involvement of different intracellular pathways in the regulation of cell proliferation appropriate activators or inhibitors
were used. Results: Hydroxylated estrogens, as E2, exhibited no influence on cAMP accumulation and PKA activation.
In concomitant treatments with forskolin, cell proliferation decreased to the amt. noted under the influence of forskolin
alone. A PKA inhibitor (PKI) had no statistically significant effect on proliferation stimulated by E2 and its hydroxylated
metabolites. Phospho-ERK1/2 protein expression in cells stimulated with E2, 2-OH-E2 and 4-OH-E2 was not
significantly different from the control. However, co-treatment with both PD98059 and E2 or its hydroxylated metabolites
reversed the effect of tested compds. on cell proliferation. Conclusions: We have shown that E2 hydroxylated
metabolites do not activate cAMP/PKA in breast cancer cells and confirm previously published data, which showed a lack
of ERK1/2 activation in a breast cancer cell line. The obsd. reversible action of PD98059 on cell proliferation can be
explained by the fact that hydroxylated estrogens, as E2, stimulate secretion of a no. of growth factors, which affect
MAPK activity, suggested by Lobenhofer et al. (2000).
~0 Citings
178. LGR6 is a high affinity receptor of R-spondins and potentially functions as a tumor suppressor
By Gong, Xing; Carmon, Kendra S.; Lin, Qiushi; Thomas, Anthony; Yi, Jing; Liu, Qingyun
Background: LGR6 (leucine-rich repeat contg., G protein-coupled receptor 6) is a member of the rhodopsin-like seven
transmembrane domain receptor superfamily with the highest homol. to LGR4 and LGR5. LGR6 was found as one of the
novel genes mutated in colon cancer through total exon sequencing and its promoter region is hypermethylated in 20-50
% of colon cancer cases. In the skin, LGR6 marks a population of stem cells that can give rise to all cell lineages.
Recently, we and others demonstrated that LGR4 and LGR5 function as receptors of R-spondins to potentiate Wnt/β-
catenin signaling. However, the binding affinity and functional response of LGR6 to R-spondins, and the activity of colon
cancer mutants of LGR6 have not been detd. Principal Findings: We found that LGR6 also binds and responds to R-
spondins 1-3 with high affinity to enhance Wnt/β-catenin signaling through increased LRP6 phosphorylation. Similar to
LGR4 and LGR5, LGR6 is not coupled to heterotrimeric G proteins or to β-arrestin following R-spondin stimulation.
Functional and expression anal. of three somatic mutations identified in colon cancer samples indicates that one mutant
fails to bind and respond to R-spondin (loss-of-function), but the other two have no significant effect on receptor function.
Overexpression of wild-type LGR6 in HeLa cells leads to increased cell migration following co-treatment with R-spondin1
and Wnt3a when compared to vector control cells or cells overexpressing the loss-of-function mutant. Conclusions:
LGR6 is a high affinity receptor for R-spondins 1-3 and potentially functions as a tumor suppressor despite its pos. effect
on Wnt/β-catenin signaling.
~20 Citings
179. Generation of neural stem cells from human trophoblast stem cells
By Lee, Jau-Nan; Lee, Tony Tung-Ying; Lee, Yuta; Tsai, Eing-Mei
SciFinder® Page 73
Provided herein are isolated neural stem cells. Also provided
are methods for treatment of neurodegenerative diseases using
suitable prepns. comprising the isolated neural stem cells.
~1 Citing
180. The cAMP signaling system inhibits the repair of γ-ray-induced DNA damage by promoting Epac1-mediated
proteasomal degradation of XRCC1 protein in human lung cancer cells
By Cho, Eun-Ah; Juhnn, Yong-Sung
CAPLUS, DOI:10.1016/j.bbrc.2012.04.139
CAMP is involved in the regulation of metab., gene expression, cellular growth and proliferation. Recently, the cAMP
signaling system was found to modulate DNA-damaging agent-induced apoptosis by regulating the expression of Bcl-2
family proteins and inhibitors of apoptosis. Thus, we hypothesized that the cAMP signaling may modulate DNA repair
activity, and we investigated the effects of the cAMP signaling system on γ-ray-induced DNA damage repair in lung
cancer cells. Transient expression of a constitutively active mutant of stimulatory G protein (GαsQL) or treatment with
forskolin, an adenylyl cyclase activator, augmented radiation-induced DNA damage and inhibited repair of the damage in
H1299 lung cancer cells. Expression of GαsQL or treatment with forskolin or isoproterenol inhibited the radiation-
induced expression of the XRCC1 protein, and exogenous expression of XRCC1 abolished the DNA repair-inhibiting
effect of forskolin. Forskolin treatment promoted the ubiquitin and proteasome-dependent degrdn. of the XRCC1 protein,
resulting in a significant decrease in the half-life of the protein after γ-ray irradn. The effect of forskolin on XRCC1
expression was not inhibited by PKA inhibitor, but 8-pCPT-2'-O-Me-cAMP, an Epac-selective cAMP analog, increased
ubiquitination of XRCC1 protein and decreased XRCC1 expression. Knockdown of Epac1 abolished the effect of 8-
pCPT-2'-O-Me-cAMP and restored XRCC1 protein level following γ-ray irradn. From these results, we conclude that the
cAMP signaling system inhibits the repair of γ-ray-induced DNA damage by promoting the ubiquitin-proteasome
dependent degrdn. of XRCC1 in an Epac-dependent pathway in lung cancer cells.
~7 Citings
181. Melanocortin 2 receptor-associated protein (MRAP) and MRAP2 in human adrenocortical tissues: regulation
of expression and association with ACTH responsiveness
By Hofland, Johannes; Delhanty, Patric J.; Steenbergen, Jacobie; Hofland, Leo J.; van Koetsveld, Peter M.; van
Nederveen, Francien H.; de Herder, Wouter W.; Feelders, Richard A.; de Jong, Frank H.
CAPLUS, DOI:10.1210/jc.2011-2328
SciFinder® Page 74
Context: ACTH stimulates adrenocortical steroid prodn. through the melanocortin 2 receptor (MC2R). MC2R trafficking
and signaling are dependent on the MC2R accessory protein (MRAP). The MRAP homolog MRAP2 also transports the
MC2R to the cell surface but might prevent activation. Objective: The objective of the investigation was to study the
regulatory pathways of MRAP and MRAP2 and their contributions to ACTH responsiveness in human adrenal tissues.
Design and Setting: MRAP, MRAP2, and MC2R expression levels were studied in 32 human adrenocortical samples.
Regulation of these mRNAs was investigated in 43 primary adrenal cultures, stimulated with ACTH, forskolin, angiotensin
II (AngII), phorbol-12-myristate-13-acetate (PMA), or dexamethasone. The induction of cortisol, cAMP, and ACTH-
responsive genes after treatment with ACTH was related to MRAP, MRAP2, and MC2R expression levels. Results:
MRAP and MRAP2 levels were lower in adrenocortical carcinomas (ACC) than in other adrenal tissues (P < 0.001).
Patient ACTH and cortisol levels were assocd. with adrenal levels of MRAP and MC2R in adrenal hyperplasia samples
(P < 0.05) but not in tumors. ACTH induced the expression of MRAP 11 ± 2.1-fold and MC2R 20 ± 3.8-fold in all adrenal
tissue types (mean ± SEM, both P < 0.0001), whereas AngII augmented these mRNAs 4.0 ± 1.2-fold and 12.6 ± 3.2-fold
(P < 0.0001) in all but the ACC. MRAP2 expression was suppressed by forskolin (-24 ± 15%, P = 0.013) and PMA (-22 ±
7%, P = 0.0007). MRAP, MRAP2, or MC2R levels were not assocd. with the induction of cortisol, cAMP, or gene
expression by ACTH in vitro. Conclusion: MRAP and MC2R expression is induced by ACTH and AngII, which would
facilitate cell surface receptor availability. Physiol. expression levels of MRAP, MRAP2, and MC2R were not limiting for
ACTH sensitivity.
~7 Citings
182. Screening of anti-cancer agent using zebrafish: Comparison with the MTT assay
By Li, Yigen; Huang, Wenjin; Huang, Shenyuan; Du, Jiulin; Huang, Cheng
CAPLUS, DOI:10.1016/j.bbrc.2012.04.110
The MTT (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyltetrazolium bromide) assay is a classical method for screening
cytotoxic anti-cancer agents. Candidate drugs from the MTT assay need in vivo models to test their efficiency and to
assess the absorption, distribution, metab., excretion, and toxicity of the drugs. An in vivo screening model could
increase the rate of development of anti-cancer drugs. Here, we used zebrafish to screen a library of 502 natural
compds. and compared the results with those from an MTT assay of the MCF7 breast cancer cell line. We identified 59
toxic compds. in the zebrafish screen, 21 of which were also identified by the MTT assay, and 28 of which were already
known for their anti-cancer and apoptosis-inducing effects. These compds. induced apoptosis and activated the p53
pathway in zebrafish within 3 h treatment. Our results indicate that zebrafish is a simple, reliable and highly efficient in
vivo tool for cancer drug screening, and could complement the MTT assay.
~28 Citings
183. Hypoxia and MITF control metastatic behavior in mouse and human melanoma cells
By Cheli, Y.; Giuliano, S.; Fenouille, N.; Allegra, M.; Hofman, V.; Hofman, P.; Bahadoran, P.; Lacour, J.-P.; Tartare-
Deckert, S.; Bertolotto, C.; et al
Melanomas are very aggressive neoplasms with notorious resistance to therapeutics. It was recently proposed that the
remarkable phenotypic plasticity of melanoma cells allows for the rapid development of both resistance to
chemotherapeutic drugs and invasive properties. Indeed, the capacity of melanoma cells to form distant metastases is
the main cause of mortality in melanoma patients. Therefore, the identification of the mechanism controlling melanoma
phenotype is of paramount importance. In the present report, we show that deletion of microphthalmia-assocd.
transcription factor (MITF), the master gene in melanocyte differentiation, is sufficient to increase the metastatic potential
of mouse and human melanoma cells. MITF silencing also increases fibronectin and Snail, two mesenchymal markers
that might explain the increased invasiveness in vitro and in vivo. Furthermore, ablation of this population by Forskolin-
induced differentiation or MITF-forced expression significantly decreases tumor and metastasis formation, suggesting
that eradication of low-MITF cells might improve melanoma treatment. Moreover, we demonstrate that a hypoxic
microenvironment decreases MITF expression through an indirect, hypoxia-inducible factor 1 (HIF1)α-dependant
transcriptional mechanism, and increases the tumorigenic and metastatic properties of melanoma cells. We identified
Bhlhb2, a new factor in melanoma biol., as the mediator of hypoxia/HIF1α inhibitory effect on MITF expression. Our
results reveal a hypoxia-HIF1α-BHLHB2-MITF cascade controlling the phenotypic plasticity in melanoma cells and
favoring metastasis development. Targeting this pathway might be helpful in the design of new anti-melanoma therapies.
~43 Citings

SciFinder® Page 75
184. Coleus forskohlii
By Kaur, Manpreet; Kaur, Harinder
From Indian Pharmacist (New Delhi, India) (2012), 10(8), 29-32. Language: English, Database: CAPLUS
A review. Coleus forskohlii [Willd] Briq., belonging to the family Labiatae (Lamiaceae), is reported to possess anti-
asthmatic, tonic, blood purifying, expectorant, anthelmintic, laxative and antispasmodic properties. It is also reported to
be used in veterinary science. Coleus forskohlii is an important source of chems. of immense medicinal and
pharmaceutical importance such as forskolin and its derivs. are effective in glaucoma, cardiovascular diseases, asthma,
allergies, psoriasis, cancer, obesity and depression. Hence, in view of the immense medicinal importance of the plant,
this review is, therefore, an effort to compile all the information reported on its phytochem. and pharmacol. activities so
that more interest could be diverted towards this herb, due to its low awareness, for the treatment and relief from various
ailments and diseases.
~0 Citings
185. Mechanisms of action of zinc on rat intestinal epithelial electrogenic ion secretion: insights into its
antidiarrhoeal actions
By Bzik, Victoria A.; Medani, Mekki; Baird, Alan W.; Winter, Desmond C.; Brayden, David J.
From Journal of Pharmacy and Pharmacology (2012), 64(5), 644-653. Language: English, Database: CAPLUS,
DOI:10.1111/j.2042-7158.2011.01441.x
Zinc is a useful addn. to oral rehydration therapy for acute diarrhoea. We have assessed the mechanism of its epithelial
antisecretory action when intestinal epithelial tight junctions were pharmacol. opened. Rat isolated ileal and colonic
mucosae were mounted in Ussing chambers and exposed to ZnSO4 (Zn2+) in the presence of secretagogues and
inhibition of short circuit current (Isc) was measured. Pre-incubation with basolateral but not apical Zn2+ reduced Isc
stimulated by forskolin, carbachol and A23187. In the presence of the tight junction-opener, cytochalasin D,
antisecretory effects of apically-applied Zn2+ were enabled in colon and ileum. The apparent permeability coeff. (Papp) of
Zn2+ was increased 1.4- and 2.4-fold across rat ileum and colon, resp., by cytochalasin D. Basolateral addn. of Zn2+ also
reduced the Isc stimulated by nystatin in rat colon, confirming K channel inhibition. In comparison with other inhibitors,
Zn2+ was a relatively weak blocker of basolateral KATP and KCa2+ channels. Exposure of ileum and colon to Zn2+ for 60
min had minimal effects on epithelial histol. Antisecretory effects of Zn2+ on intestinal epithelia arose in part through
nonselective blockade of basolateral K channels, which was enabled when tight junctions were open.
~5 Citings
186. Suppression of adipose lipolysis by long-chain fatty acid analogs

By Kalderon, Bella; Azazmeh, Narmen; Azulay, Nili; Vissler, Noam; Valitsky, Michael; Bar-Tana, Jacob
From Journal of Lipid Research (2012), 53(5), 868-878. Language: English, Database: CAPLUS,
DOI:10.1194/jlr.M022673
Agonist-induced lipolysis of adipose fat is robustly inhibited by insulin or by feedback inhibition by the long-chain fatty
acids (LCFA) produced during lipolysis. However, the mode of action of LCFA in suppressing adipose lipolysis is not
clear. β,β'-Tetra-Me hexadecanedioic acid (Mββ/ EDICA16) is a synthetic LCFA that is neither esterified into lipids nor β-
oxidized, and therefore, it was exploited for suppressing agonist-induced lipolysis in analogy to natural LCFA. Mββ is
shown here to suppress isoproterenol-induced lipolysis in the rat in vivo as well as in 3T3-L1 adipocytes. Inhibition of
isoproterenol-induced lipolysis is due to decrease in isoproterenol-induced cAMP with concomitant inhibition of the
phosphorylation of hormone-sensitive lipase and perilipin by protein kinase A. Suppression of cellular cAMP levels is
accounted for by inhibition of the adenylate cyclase due to suppression of Raf1 expression by Mββ-activated AMPK.
Suppression of Raf1 is further complemented by induction of components of the unfolded-protein-response by Mββ. Our
findings imply genuine inhibition of agonist-induced adipose lipolysis by LCFA, independent of their β-oxidn. or
reesterification. Mββ suppression of agonist-induced lipolysis and cellular cAMP levels independent of the insulin
transduction pathway may indicate that synthetic LCFA could serve as insulin mimetics in the lipolysis context under
conditions of insulin resistance.
~4 Citings
187. Potassium channel mutant KCNJ5 T158A expression in HAC-15 cells increases aldosterone synthesis
By Oki, Kenji; Plonczynski, Maria W.; Lam, Milay Luis; Gomez-Sanchez, Elise P.; Gomez-Sanchez, Celso E.
SciFinder® Page 76
Primary aldosteronism is the most common cause of secondary hypertension, most frequently due to an aldosterone-
producing adenoma or idiopathic hyperaldosteronism. Somatic mutations of the potassium channel KCNJ5 in the region
of the selectivity filter have been found in a significant no. of aldosterone-producing adenomas. There are also familial
forms of primary aldosteronism, one of which, familial hyperaldosteronism type 3 which to date has been found in one
family who presented with a severe abnormality in aldosterone and 18-oxocortisol prodn. and hypertrophy and
hyperplasia of the transitional zone of the adrenal cortex. In familial hyperaldosteronism type 3, there is a genomic
mutation causing a T158A change of amino acids within the selectivity filter region of the KCNJ5 gene. We are reporting
our studies demonstrating that lentiviral-mediated expression of a gene carrying the T158A mutation of the KCNJ5 in the
HAC15 adrenal cortical carcinoma cell line causes a 5.3-fold increase in aldosterone secretion in unstimulated HAC15-
KCNJ5 cells and that forskolin-stimulated aldosterone secretion was greater than that of angiotensin II. Expression of
the mutated KCNJ5 gene decreases plasma membrane polarization, allowing sodium and calcium influx into the cells.
The calcium channel antagonist nifedipine and the calmodulin inhibitor W-7 variably inhibited the effect. Overexpression
of the mutated KCNJ5 channel resulted in a modest decrease in HAC15 cell proliferation. These studies demonstrate
that the T158A mutation of the KCNJ5 gene produces a marked stimulation in aldosterone biosynthesis that is dependent
on membrane depolarization and sodium and calcium influx into the HAC15 adrenal cortical carcinoma cells.
~58 Citings
188. Primary aldosteronism takes (KCNJ)five!

By Zennaro, Maria-Christina
A review. The research of Oki et al. (2012), entitled 'Potassium channel mutant KCNJ5 T158A expression in HAC-15
cells increases aldosterone synthesis' is reviewed with commentary and refs. These authors demonstrate that the
inherited KCNJS T158A mutation produces a marked stimulation in aldosterone biosynthesis that is dependent on
membrane depolarization followed by calcium influx into adrenal cortical carcinoma cells. Oki et al. tackles a central
issue that had not previously been addressed, which is the causal link between KCNJ5 mutations, membrane
depolarization, aldosterone overprodn., and cell proliferation. The authors show that expression of channels harboring
the T158A mutation potentiated basal aldosterone prodn., which was further stimulated by angiotensin II and the protein
kinase A activator forskolin.
~1 Citing
189. The gene SLC41A1 transport protein as a Na+/Mg2+ exchanger and biological functions of the transporter
By Schweigel, Monika; Kolisek, Martin
From Eur. Pat. Appl. (2012), EP 2431386 A1 20120321, Language: English, Database: CAPLUS
The SLC41A1 gene is shown to encode a sodium/magnesium
exchanger and the function of the exchanger is examd. at the
cell, tissue, organ and organism level. Libraries of mutant
alleles of the SLC41A1 gene, and antibodies to the gene
products that were used in identifying its function are
described. Methods of screening for effectors of the protein,
e.g. for therapeutic use, are described. In part, it is related to
methods useful in (a) identifying mols. that bind SLC41A1
polypeptides, which (b) modulate SLC41A1 related Na+/Mg2+
exchanger activity or its cellular or tissue specific expression.
Thus, the invention comprises SLC41A1 mutation libraries,
SLC41A1 specific antibodies, their generation and finding as
well as an inducible conditional SLC41A1 knock out mice
model and inducible conditional knock out SLC41A1 cell lines.
~0 Citings
190. Methods for identifying gene SLC41A1-encoded Na+/Mg2+ exchanger and functions of transporter
By Schweigel, Monika; Kolisek, Martin
SciFinder® Page 77
The invention relates to methods to identify gene SLC41A1-
encoded sodium/magnesium exchanger that functions at the
cell, tissue, organ and organism level. In part, it is related to
methods useful in identifying mols. that bind SLC41A1
polypeptides, which modulate SLC41A1 related Na+/Mg2+
exchanger activity or its cellular or tissue specific expression.
Thus, the invention comprises SLC41A1 mutation libraries,
SLC41A1 specific antibodies, their generation and finding as
well as an inducible conditional SLC41A1 knock out mice
model and inducible conditional knock out SLC41A1 cell lines.
Methods of screening for effectors of the protein, e.g. for
therapeutic use, are described.
~0 Citings
191. Oncogenic K-ras expression is associated with derangement of the cAMP/PKA pathway and forskolin-
reversible alterations of mitochondrial dynamics and respiration
By Palorini, R.; De Rasmo, D.; Gaviraghi, M.; Danna, L. Sala; Signorile, A.; Cirulli, C.; Chiaradonna, F.; Alberghina, L.;
Papa, S.
The Warburg effect in cancer cells has been proposed to involve several mechanisms, including adaptation to hypoxia,
oncogenes activation or loss of onco-suppressors and impaired mitochondrial function. In previous papers, it has been
shown that K-ras transformed mouse cells are much more sensitive as compared with normal cells to glucose withdrawal
(undergoing apoptosis) and present a high glycolytic rate and a strong redn. of mitochondrial complex I. Recent
observations suggest that transformed cells have a derangement in the cAMP/cAMP-dependent protein kinase
(cAMP/PKA) pathway, which is known to regulate several mitochondrial functions. Herein, the derangement of the
cAMP/PKA pathway and its impact on transformation-linked changes of mitochondrial functions is investigated.
Exogenous stimulation of PKA activity, achieved by forskolin treatment, protected K-ras-transformed cells from apoptosis
induced by glucose deprivation, enhanced complex I activity, intracellular ATP levels, mitochondrial fusion and
decreased intracellular reactive oxygen species (ROS) levels. Several of these effects were almost completely
prevented by inhibiting the PKA activity. Short-time treatment with compds. favoring mitochondrial fusion strongly
decreased the cellular ROS levels esp. in transformed cells. These findings support the notion that glucose shortage-
induced apoptosis, specific of K-ras-transformed cells, is assocd. to a derangement of PKA signaling that leads to
mitochondrial complex I decrease, redn. of ATP formation, prevalence of mitochondrial fission over fusion, and thereby
opening new approaches for development of anticancer drugs.
~14 Citings
192. Traditional chinese medicine composition for resisting tumor

By Fan, Shenggang; Liu, Peng
The title traditional Chinese medicine composition is manufactured from (by wt. parts): Astragalus membranaceus 5-15,
Panax ginseng 10-20, Coleus forskolin 5-10, Ganoderma 5-15, Scutellaria barbata 10-25, Hedyotis diffusa 5-25, and
Citrus reticulata 5-10. The traditional Chinese medicine composition can be used for resisting liver cancer, gastric
cancer, renal cancer, oesophagus cancer, lung cancer and leukemia.
~0 Citings
193. Role of Epac and protein kinase A in thyrotropin-induced gene expression in primary thyrocytes
By van Staveren, Wilma C. G.; Beeckman, Sandrine; Tomas, Gil; Dom, Genevieve; Hebrant, Aline; Delys, Laurent;
Vliem, Marjolein J.; Tresallet, Christophe; Andry, Guy; Franc, Brigitte; et al
DOI:10.1016/j.yexcr.2011.12.022
SciFinder® Page 78
CAMP pathway activation by TSH induces differentiation and gene expression in thyrocytes. We investigated which
partners of the cAMP cascade regulate gene expression modulations: protein kinase A and/or the exchange proteins
directly activated by cAMP (Epac). Human primary cultured thyrocytes were analyzed by microarrays after treatment
with the adenylate cyclase activator forskolin, the protein kinase A (PKA) activator 6-MB-cAMP and the Epac-selective
cAMP analog 8-pCPT-2'-O-Me-cAMP (007) alone or combined with 6-MB-cAMP. Profiles were compared to those of
TSH. Cultures treated with the adenylate cyclase- or the PKA activator alone or the latter combined with 007 had profiles
similar to those induced by TSH. mRNA profiles of 007-treated cultures were highly distinct from TSH-treated cells,
suggesting that TSH-modulated gene expressions are mainly modulated by cAMP and PKA and not through Epac in
cultured human thyroid cells. To investigate whether the Epac-Rap-RapGAP pathway could play a potential role in
thyroid tumorigenesis, the mRNA expressions of its constituent proteins were investigated in two malignant thyroid tumor
types. Modulations of this pathway suggest an increased Rap pathway activity in these cancers independent from cAMP
activation.
~4 Citings
194. MicroRNA 335 is required for differentiation of malignant glioma cells induced by activation of
cAMP/protein kinase A pathway
By Shu, Minfeng; Zhou, Yuehan; Zhu, Wenbo; Zhang, Haipeng; Wu, Sihan; Chen, Jingkao; Yan, Guangmei
DOI:10.1124/mol.111.076166
Glioma is the most common malignant cancer affecting the central nerve system, with dismal prognosis. Differentiation-
inducing therapy is a novel strategy that has been preliminarily proved effective against malignant glioma. We have
reported previously that activation of cAMP/protein kinase A (PKA) pathway is capable of inducing glioma cell
differentiation, characterized by astrocyte-like shape and dramatic induction of astrocyte biomarker glial fibrillary acidic
protein (GFAP). However, little progress has been made on mol. mechanisms related. Here we demonstrate that
microRNA 335 (miR-335) is responsible for the glioma cell differentiation stimulated by activation of cAMP/PKA pathway.
In the cAMP elevator cholera toxin-induced differentiation model of rat C6 glioma cells, miR-335 was significantly up-
regulated, which was mimicked by other typical cAMP/PKA pathway activators (e.g., forskolin, dibutyryl-cAMP) and
abolished by PKA-specific inhibitor (9R,10S,12S)-2,3,9,10,11,12-hexahydro-10-hydroxy-9-methyl-1-oxo-9,12-epoxy-1H-
diindolo[1,2,3-fg:3',2',1'-kl]pyrrolo[3,4-i][1,6]benzodiazocine-10-carboxylic acid, hexyl ester (KT5720). In an assay
measuring gain and loss of miR-335 function, exogenetic miR-335 resulted in induction of GFAP, whereas miR-335
specific inhibitor antagomir-335 violently blocked cholera toxin-induced GFAP up-regulation. It is noteworthy that in
human U87-MG glioma cells and human primary culture glioma cells, miR-335 also mediated cholera toxin-induced
differentiation. Taken together, our findings suggest that miR-335 is potently required for differentiation of malignant
glioma cells induced by cAMP/PKA pathway activation, and a single microRNA may act as an important fate determinant
to control the differentiation status of malignant gliomas, which has provided a new insight into differentiation-inducing
therapy against malignant gliomas.
~19 Citings
195. Cobinamides are novel coactivators of nitric oxide receptor that target soluble guanylyl cyclase catalytic
domain
By Sharina, Iraida; Sobolevsky, Michael; Doursout, Marie-Francoise; Gryko, Dorota; Martin, Emil
From Journal of Pharmacology and Experimental Therapeutics (2012), 340(3), 723-732. Language: English, Database:
CAPLUS, DOI:10.1124/jpet.111.186957
Sol. guanylyl cyclase (sGC), a ubiquitously expressed heme-contg. receptor for nitric oxide (NO), is a key mediator of
NO-dependent processes. In addn. to NO, a no. of synthetic compds. that target the heme-binding region of sGC and
activate it in a NO-independent fashion have been described. We report here that dicyanocobinamide (CN2-Cbi), a
naturally occurring intermediate of vitamin B12 synthesis, acts as a sGC coactivator both in vitro and in intact cells. Heme
depletion or heme oxidn. does not affect CN2-Cbi-dependent activation. Deletion mutagenesis demonstrates that CN2-
Cbi targets a new regulatory site and functions though a novel mechanism of sGC activation. Unlike all known sGC
regulators that target the N-terminal regulatory regions, CN2-Cbi directly targets the catalytic domain of sGC, resembling
the effect of forskolin on adenylyl cyclases. CN2-Cbi synergistically enhances sGC activation by NO-independent
regulators 3-(4-amino-5-cyclopropylpyrimidine-2-yl)-1-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyridine (BAY41-2272), 4-[((4-
carboxybutyl){2-[(4-phenethylbenzyl)oxy]phenethyl}amino) Me [benzoic]-acid (cinaciguat or BAY58-2667), and 5-chloro-
2-(5-chloro-thiophene-2-sulfonylamino-N-(4-(morpholine-4-sulfonyl)-phenyl)-benzamide sodium salt (ataciguat or HMR-
1766). BAY41-2272 and CN2-Cbi act reciprocally by decreasing the EC50 values. CN2-Cbi increases intracellular cGMP
levels and displays vasorelaxing activity in phenylephrine-constricted rat aortic rings in an endothelium-independent
manner. Both effects are synergistically potentiated by BAY41-2272. These studies uncover a new mode of sGC
regulation and provide a new tool for understanding the mechanism of sGC activation and function. CN2-Cbi also offers
new possibilities for its therapeutic applications in augmenting the effect of other sGC-targeting drugs.
SciFinder® Page 79
~14 Citings
196. In vitro propagation studies on Coleus forskohlii Briq. - a medicinal plant

By Praveena, R.; Amarnath Pandian, S.; Jegadeesan, M.
From International Journal of Pharma and Bio Sciences (2012), 3(1), 82-92. Language: English, Database: CAPLUS
Coleus forskohlii Briq. (Lamiaceae) is an important medicinal plant with excellent export potential in herbal drug trade.
The root of this plant is medicinally useful for high blood pressure, spasmolysis, obesity and constipation. A diterpene
compd., forskolin, obtained from tubers root of this plant is used for glaucoma congestive, asthma and certain cancers.
The present work aims at evolving a protocol for rapid multiplication using culture technique. Tissue explants from leaf
lamina, leaf base and node were cultured on MS medium supplemented with different concns. (0.5-2.0 mg/l) of IAA, 2, 4-
D and BAP and their growth responses like callusing and shooting and rooting were elucidated. It was obsd. that leaf
base explants elicited max. callusing, shooting and rooting response than that of node and leaf lamina. Multiple shoots
were obtained from leaf base explants. This indicates the possibility of using tissue culture protocol for com. scale of
prodn.
~1 Citing
197. Methylseleninic acid is a novel suppressor of aromatase expression

By Gao, Ruijuan; Zhao, Lijuan; Liu, Xichun; Rowan, Brian G.; Wabitsch, Martin; Edwards, Dean P.; Nishi, Yoshihiro;
Yanase, Toshihiko; Yu, Qun; Dong, Yan
From Journal of Endocrinology (2012), 212(2), 199-205. Language: English, Database: CAPLUS, DOI:10.1530/JOE-
11-0363
Elevated circulating estrogen levels, as a result of increased peripheral aromatization of androgens by aromatase, have
been indicated to underlie the assocn. between obesity and a higher risk of breast cancer in postmenopausal women.
Although aromatase inhibitors have been used as a first-line therapy for estrogen receptor-pos. breast cancer in
postmenopausal women, their potential as breast cancer chemopreventive agents has been limited due to toxicities and
high costs. It is therefore imperative to develop new aromatase-inhibiting/suppressing agents with lower toxicities and
lower costs for breast cancer chemoprevention, esp. in obese postmenopausal women. The expression of the
aromatase gene, CYP19, is controlled in a tissue-specific manner by the alternate use of different promoters. In obese
postmenopausal women, increased peripheral aromatase is primarily attributed to the activity of the glucocorticoid-
stimulated promoter, PI.4, and the cAMP-stimulated promoter, PII. In the present study, we show that methylseleninic
acid (MSA), a second-generation selenium compd., can effectively suppress aromatase activation by dexamethasone, a
synthetic glucocorticoid, and forskolin, a specific activator of adenylate cyclase. Unlike the action of aromatase inhibitors,
MSA suppression of aromatase activation is not mediated via direct inhibition of aromatase enzymic activity. Rather, it is
attributable to a marked downregulation of promoters PI.4- and PII-specific aromatase mRNA expression, and thereby a
redn. of aromatase protein. Considering the low-cost and low-toxicity nature of MSA, our findings provide a strong
rationale for the further development of MSA as a breast cancer chemopreventive agent for obese postmenopausal
women.
~3 Citings
198. The influence of Aspalathus linearis (Rooibos) and dihydrochalcones on adrenal steroidogenesis:
Quantification of steroid intermediates and end products in H295R cells
By Schloms, Lindie; Storbeck, Karl-Heinz; Swart, Pieter; Gelderblom, Wentzel C. A.; Swart, Amanda C.
From Journal of Steroid Biochemistry and Molecular Biology (2012), 128(3-5), 128-138. Language: English, Database:
SciFinder® Page 80
The steroid hormone output of the adrenal gland is crucial in the maintenance of hormonal homeostasis, with hormonal
imbalances being assocd. with numerous clin. conditions which include, amongst others, hypertension, metabolic
syndrome, cardiovascular disease, insulin resistance and type 2 diabetes. Aspalathus linearis (Rooibos), which has
been reported to aid stress-related symptoms linked to metabolic diseases, contains a wide spectrum of bioactive
phenolic compds. of which Aspalathin is unique. In this study the inhibitory effects of Rooibos and the dihydrochalcones,
Aspalathin and Nothofagin, were investigated on adrenal steroidogenesis. The activities of both cytochrome P 450 17α-
hydroxylase/17,20 lyase and cytochrome P 450 21-hydroxylase were significantly inhibited in COS-1 cells. In order to
study the effect of these compds. in H295R cells, a human adrenal carcinoma cell line, a novel UPLC-MS/MS method
was developed for the detection and quantification of twenty-one steroid metabolites using a single chromatog. sepn.
Under both basal and forskolin-stimulated conditions, the total amt. of steroids produced in H295R cells significantly
decreased in the presence of Rooibos, Aspalathin and Nothofagin. Under stimulated conditions, Rooibos decreased the
total steroid output 4-fold and resulted in a significant redn. of aldosterone and cortisol precursors.
Dehydroepiandrosterone-sulfate levels were unchanged, while the levels of androstenedione (A4) and 11β-
hydroxyandrostenedione (11βOH-A4) were inhibited 5.5 and 2.3-fold, resp. Quantification of 11βOH-A4 showed this
metabolite to be a major product of steroidogenesis in H295R cells and we confirm, for the first time, that this steroid
metabolite is the product of the hydroxylation of A4 by human cytochrome P 450 11β-hydroxylase. Taken together our
results demonstrate that Rooibos, Aspalathin and Nothofagin influence steroid hormone biosynthesis and the flux
through the mineralocorticoid, glucocorticoid and androgen pathways, thus possibly contributing to the alleviation of neg.
effects arising from elevated glucocorticoid levels.
~28 Citings
199. Activation of LTRs from different human endogenous retrovirus (HERV) families by the HTLV-1 Tax protein
and T-cell activators
By Toufaily, Chirine; Landry, Sebastien; Leib-Mosch, Christine; Rassart, Eric; Barbeau, Benoit
From Viruses (2011), 3, 2146-2159. Language: English, Database: CAPLUS, DOI:10.3390/v3112146
Human endogenous retroviruses (HERVs) represent approx. 8% of our genome. HERVs influence cellular gene
expression and contribute to normal physiol. processes such as cellular differentiation and morphogenesis. HERVs have
also been assocd. with certain pathol. conditions, including cancer and neurodegenerative diseases. As HTLV-1 causes
adult T-cell leukemia and HTLV-1-assocd. myelopathy/tropical spastic paraparesis (HAM/TSP) and has been shown to
modulate host gene expression mainly through the expression of the powerful Tax transactivator, herein we were
interested in looking at the potential modulation capacity of HTLV-1 Tax on HERV expression. In order to evaluate the
promoter activity of different HERV LTRs, pHERV-LTR-luc constructs were co-transfected in Jurkat T-cells with a Tax
expression vector. Tax expression potently increased the LTR activity of HERV-W8 and HERV-H (MC16). In parallel,
Jurkat cells were also stimulated with different T-cell-activating agents and HERV LTRs were obsd. to respond to
different combination of Forskolin, bpV[pic] a protein tyrosine phosphatase inhibitor, and PMA. Transfection of
expression vectors for different Tax mutants in Jurkat cells showed that several transcription factors including CREB
appeared to be important for HERV-W8 LTR activation. Deletion mutants were derived from the HERV-W8 LTR and the
region from -137 to -123 was found to be important for LTR response following Tax expression in Jurkat cells, while a
different region was shown to be required in cells treated with activators. Our results thus demonstrated that HTLV-1 Tax
activates several HERV LTRs. This raises the possibility that upregulated HERV expression could be involved in
diseases assocd. with HTLV-1 infection.
~13 Citings
200. Rimonabant, a cannabinoid receptor type 1 inverse agonist, inhibits hepatocyte lipogenesis by activating
liver kinase B1 and AMP-activated protein kinase axis downstream of Gαi/o inhibition
By Wu, Hong Min; Yang, Yoon Mee; Kim, Sang Geon
DOI:10.1124/mol.111.072769
SciFinder® Page 81
Liver X receptor-α (LXRα) and its target sterol regulatory element-binding protein-1c (SREBP-1c) play key roles in
hepatic lipogenesis. Rimonabant, an inverse agonist of cannabinoid receptor type 1 (CB1), has been studied as an
antiobesity drug. In view of the link between CB1 and energy metab., this study investigated the effect of rimonabant on
LXRα-mediated lipogenesis in hepatocytes and the underlying basis. Rimonabant treatment inhibited CYP7A1-LXRα
response element gene transactivation and an increase in LXRα mRNA level by the LXRα agonist N-(2,2,2-
trifluoroethyl)-N-[4-[2,2,2-trifluoro-1-hydroxy-1-(trifluoromethyl)ethyl]phenyl]-benzenesulfonamide (T0901317) in HepG2
cells. Rimonabant consistently attenuated the activation of SREBP-1c and its target gene induction. The reversal by
CB1 agonists on rimonabant's repression of SREBP-1c supported the role of CB1 in this effect. Rimonabant inhibited
the activation of SREBP-1c presumably via Gαi/o inhibition, as did pertussis toxin. Adenylyl cyclase activator forskolin or
8-bromo-cAMP treatment mimicked the action of rimonabant, suggesting that Gαi/o inhibition causes repression of
SREBP-1c by increasing the cAMP level. Knockdown or chem. inhibition of protein kinase A (PKA) prevented the
inhibition of LXRα by rimonabant, supporting the fact that an increase in cAMP content and PKA activation, which
catalyzes LXRα inhibitory phosphorylation, might be responsible for the antilipogenic effect. In addn., rimonabant
activated liver kinase B1 (LKB1), resulting in the activation of AMP-activated protein kinase responsible for LXRα
repression. Moreover, PKA inhibition prevented the activation of LKB1, supporting the fact that PKA regulates LKB1. In
conclusion, rimonabant has an antilipogenic effect in hepatocytes by inhibiting LXRα-dependent SREBP-1c induction, as
mediated by an increase in PKA activity and PKA-mediated LKB1 activation downstream of CB1-coupled Gαi/o inhibition.
~24 Citings
201. β-Adrenergic receptors (β-AR) regulate VEGF and IL-6 production by divergent pathways in high β-AR-
expressing breast cancer cell lines
By Madden, Kelley S.; Szpunar, Mercedes J.; Brown, Edward B.
DOI:10.1007/s10549-011-1348-y
Activation of β-adrenergic receptors (β-AR) drives proangiogenic factor prodn. in several types of cancers. To examine
β-AR regulation of breast cancer pathogenesis, β-AR d., signaling capacity, and functional responses to β-AR stimulation
were studied in four human breast adenocarcinoma cell lines. β-AR d. ranged from very low in MCF7 and MB-361 to
very high in MB-231 and in a brain-seeking variant of MB-231, MB-231BR. Consistent with β-AR d., β-AR activation
elevated cAMP in MCF7 and MB-361 much less than in MB-231 and MB-231BR. Functionally, β-AR stimulation did not
markedly alter vascular endothelial growth factor (VEGF) prodn. by MCF7 or MB-361. In the two high β-AR-expressing
cell lines MB-231 and MB-231BR, β-AR-induced cAMP and VEGF prodn. differed considerably, despite similar β-AR d.
The β2-AR-selective agonist terbutaline and the endogenous neurotransmitter norepinephrine decreased VEGF prodn.
by MB-231, but increased VEGF prodn. by MB-231BR. Moreover, β2-AR activation increased IL-6 prodn. by both MB-
231 and MB-231BR. These functional alterations were driven by elevated cAMP, as direct activation of adenylate
cyclase by forskolin elicited similar alterations in VEGF and IL-6 prodn. The protein kinase A antagonist KT5720
prevented β-AR-induced alterations in MB-231 and MB-231BR VEGF prodn., but not IL-6 prodn. Conclusions β-AR
expression and signaling is heterogeneous in human breast cancer cell lines. In cells with high β-AR d., β-AR stimulation
regulates VEGF prodn. through the classical β-AR-cAMP-PKA pathway, but this pathway can elicit directionally opposite
outcomes. Furthermore, in the same cells, β-AR activate a cAMP-dependent, PKA-independent pathway to increase IL-
6 prodn. The complexity of breast cancer cell β-AR expression and functional responses must be taken into account
when considering β-AR as a therapeutic target for breast cancer treatment.
~31 Citings
202. Methods for expansion and directed differentiation of human epidermal neural crest stem cells and their
use in treatment of diseases
By Sieber-Blum, Maya
SciFinder® Page 82
The invention relates to the expansion and directed differentiation of epidermal neural crest stem cells, and more
specifically human epidermal neural crest stem cells (hEPI-NCSC)into dopaminergic neurons, dopaminergic neuronal
progenitors, sensory neurons, sympathetic neurons, cholinergic neurons, melanocytes, Schwann cells, smooth muscle,
osteogenic differentiation into osteocytes, bone and cartilage and adrenergic differentiation and their use in treatment of
diseases. The hEPI-NCSC are readily accessible in the hair follicles. The patient's own hEPI-NCSC could therefore be
harvested, expanded ex vivo and then used for autologous transplantation. The present invention provides benefits over
the prior art by providing a range of effective and high yield culture methods for the differentiation of adult hEPI-NCSCs in
vitro. Such differentiated hEPI-NCSCs hold significant therapeutic potential for the treatment of various neoplastic and
non-neoplastic diseases including Parkinson's disease, multiple sclerosis, amyotrophic lateral sclerosis (ALS),
osteoarthritis, osteoporosis, stroke, and cerebellar degeneration, and various acute and severely debilitating trauma
episodes such as spinal cord injury, head and neck injury, and severe burn and/or wound repair. HEPI-NCSCs are also
very useful candidates for the study of and intervention in the aging process, and hold significant potential for use as both
genetic delivery vehicles and high throughput drug screening and/or drug discovery techniques, with the former being of
specific relevance to the treatment of neurogenetic diseases.
~1 Citing
203. Gefitinib induces apoptosis in human glioma cells by targeting Bad phosphorylation
By Chang, Cheng-Yi; Shen, Chiung-Chyi; Su, Hong-Lin; Chen, Chun-Jung
From Journal of Neuro-Oncology (2011), 105(3), 507-522. Language: English, Database: CAPLUS,
DOI:10.1007/s11060-011-0632-3
Gefitinib, a selective epidermal growth factor receptor tyrosine kinase inhibitor, is under clin. testing and use in cancer
patients, including glioma. However, the mol. mechanisms involved in gefitinib-mediated anticancer effects against
glioma remain largely uncharacterized. Gefitinib inhibits cell growth and induces apoptosis in human glioma cells.
Gefitinib also induces death of H4 cells with characteristics of the intrinsic apoptotic pathway, including Bax mitochondrial
translocation, mitochondrial outer membrane permeabilization, cytochrome c cytosolic release, and caspase-9/caspase-3
activation. The importance of Bax in mediating gefitinib-induced apoptosis was confirmed by the attenuation of apoptosis
by Bax siRNA and Bax channel blocker. Gefitinib caused Bad dephosphorylation, particularly in serine-112, and
increased its binding preference to Bcl-2 and Bcl-xL. The dephosphorylation of Bad in gefitinib-treated cells was
accompanied by reduced intracellular cAMP content and protein kinase A (PKA) activity. Adenylyl cyclase activator
forskolin attenuated, but PKA inhibitor H89 augmented, gefitinib-induced Bad dephosphorylation, Bax mitochondrial
translocation, caspase-9/caspase-3 activation, and viability loss. Intriguingly, a nonselective protein phosphatase
inhibitor okadaic acid alleviated gefitinib-induced alterations, except Bad dephosphorylation. In parallel with the higher
basal PKA activity, response of U87 cells to gefitinib treatment was delayed and relatively resistant compared with that of
H4 and T98G cells. Inactivation of PKA sensitized H4, T98G, and U87 cells to gefitinib cytotoxicity, Bad
dephosphorylation in serine-112, and caspase-9/caspase-3 activation. Our findings suggest the involvement of the
Bad/Bax signaling pathway in gefitinib-induced glioma apoptosis. Furthermore, the inactivation of PKA was shown to
play a role in triggering the proapoptotic function of Bad.
~6 Citings
204. Synthesis and metabolism of melatonin in the skin and retinal pigment epithelium
By Slominski, Radomir; Slominski, Andrzej T.
Edited By:Watson, Ronald Ross
From Melatonin in the Promotion of Health (2nd Edition) (2012), 69-80. Language: English, Database: CAPLUS,
DOI:10.1201/b11101-4
A review on melatonin in the skin, synthesis of melatonin in the skin, metab. of melatonin, and melatonin in retinal
pigmental epithelium.
~0 Citings
205. Red blood cell aggregation changes are depended on its initial value: Effect of long-term drug treatment
and short-term cell incubation with drug
By Muravyov, A. V.; Tikhomirova, I. A.; Maimistova, A. A.; Bulaeva, S. V.; Mikhailov, P. V.; Kislov, N. V.
From Clinical Hemorheology and Microcirculation (2011), 48(4), 231-240. Language: English, Database: CAPLUS,
DOI:10.3233/CH-2011-1415
SciFinder® Page 83
This study was designed to investigate whether the red cell aggregation depends on its initial level under drug therapy or
cell incubation with bioactive chem. compds. Sixty six subjects were enrolled onto this study, and sub-divided into two
groups: the first group of patients (n = 36) with cerebral atherosclerosis received pentoxifylline therapy (400 mg, thrice
daily) for 4 wk. The patients of the second group were initially treated with Epoetin beta 10,000 units s.c. thrice a week,
for 4 wk. The second group - adult anemic patients (n = 30) with the confirmed diagnosis of solid cancer (Hb < 100 g/L).
After 4 wk of pentoxifylline treatment the red cell aggregation increased (p < 0.05) in the patients with initially low RBCA.
On the other hand in the patients with initially high RBCA treatment with pentoxifylline reduced it markedly (p < 0.01). In
vitro expts. with pentoxifylline RBC incubation resulted in a decrease of the initially high RBCA by 47% (p < 0.01),
whereas in the sub-group with initially low RBCA it increased. It was obsd. that after 4 wk of epoetin-beta treatment 75%
the anemic patients with initially high RBCA had an aggregation lowering. The drop of aggregation was about 34% (p <
0.01). At the same time 25% of the study patients had a significant RBCA increase (p < 0.05) after treatment. The
initially low red cell aggregation after incubation with epoetin-beta was markedly increased by 122% (p < 0.05). On the
contrary initially high RBCA was reduced by 47% (p < 0.05). When forskolin (10 µM) was added to the RBC suspensions
the RBCA was increased in sub-group of subjects with initially low aggregation and it was decreased in sub-group with
initially high one. The similar RBCA changes were obsd. when RBC suspensions were incubated with vinpocetine,
calcium ionophore (A23187), Phorbol 12-myristate 13-acetate (PMA) as a protein kinase C (PKC) stimulator. A major
finding of this study is that the red cell aggregation effects of some drugs depend markedly on the initial, pre-treatment
aggregation status of the patients. These results demonstrate that the different red blood cell aggregation responses to
the biol. stimuli depend strongly on the initial, pre-treatment status of the subject and the most probably it is connected
with the crosstalk between the adenylyl cyclase signaling pathway and Ca2+ regulatory mechanism.
~1 Citing
206. Regulation of aromatase expression by hormones, drugs, pesticides and environmental pollutants in
canine mammary CMT-U27 cells
By Mavridis, Savvas K.; Pappas, Ioannis S.
From Epitheorese Klinikes Farmakologias kai Farmakokinetikes, International Edition (2010), 24(2), 171-174.
Language: English, Database: CAPLUS
Aromatase, the enzyme that converts testosterone to 17β-estradiol, has attracted attention lately because of its
involvement in some forms of breast cancer in women. Studies are now undergoing on a variety of species to elucidate
the effect of various agents on aromatase expression. Dog is an important species, widely used by the pharmaceutical
industry for many study types, including those that will impact decision on drug development. In the present study, a
wide variety of agents, including steroid hormones, drugs, pesticides, and environmental pollutants, were used to
investigate their effect on aromatase expression in the canine mammary CMT-U27 cell line using quant. real-time
polymerase chain reaction (qPCR) on DNA synthesized from RNA extd. from the cells.
~0 Citings
207. Molecular mechanisms of A3 adenosine receptor-induced G1 cell cycle arrest and apoptosis in androgen-
dependent and independent prostate cancer cell lines: involvement of intrinsic pathway
By Aghaei, Mahmoud; Panjehpour, Mojtaba; Karami-Tehrani, Fatemeh; Salami, Siamak
From Journal of Cancer Research and Clinical Oncology (2011), 137(10), 1511-1523. Language: English, Database:
CAPLUS, DOI:10.1007/s00432-011-1031-z
Purpose A3 adenosine receptor has shown several physiol. and pathol. activities, including cell proliferation and
apoptosis in various cancer cell lines. This study is designed to investigate mol. mechanism and apoptotic pathway of A3
adenosine receptor in DU-145, PC3 and LNcap-FGC10 human prostate cancer cells. Methods The expression level of
A3 adenosine receptor was examd. using real-time RT-PCR. cAMP concn. was also measured. MTT viability, cell
counting and BrdU incorporation tests were used to study the cell proliferation effect of IB-MECA. Cell cycle anal.,
Annexin V-FITC staining, Hoechst 33258 staining, mitochondrial membrane potential (∆ΨM), caspase-3 activity, Bcl-2
and Bax protein expression were used to detect apoptosis. Result A3 adenosine receptors mRNAs were detected at
different levels. IB-MECA inhibited forskolin-stimulated cAMP. IB-MECA at (1 µM) suppressed cell proliferation and
induced G1 cell cycle arrest. Indeed, IB-MECA down-regulated the expression of CDK4, cyclin D1 and up-regulated p53
expression. IB-MECA at (10-100 µM) induced apoptosis. The activity of caspase-3 was also increased. Expression of
Bcl-2 was decreased in response to IB-MECA, while the expression of Bax protein was increased. The results showed a
significant loss of ∆ΨM, in a dose-dependent manner. Conclusion This study introduces a possible mechanism through
A3 adenosine receptor activation. IB-MECA inhibited prostate cancer cells proliferation and induced G1 cell cycle arrest
through p53, Cdk4/cyclinD1 pathway. Apoptosis detd. by characteristic morphol. changes and increased in sub-G1
population. Loss of MMP, activation of caspase-3 and down-regulation of Bcl-2 expression indicated mitochondrial
signaling pathway that involved in the apoptosis.
~18 Citings
SciFinder® Page 84
208. Methods for the treatment of non-Hodgkin's lymphomas using lenalidomide, and gene and protein
biomarkers as a predictor
By Schafer, Peter H.; Zhang, Ling-Hua; Bartlett, J. Blake; Heise, Carla
Methods of treating or managing specific cancers, including
non-Hodgkin's lymphoma, by the administration of lenalidomide
(3-(4-amino-l-oxo-l,3-dihydro-isoindol-2-yl)- piperidine-2,6-
dione) are disclosed. Methods of using gene and protein
biomarkers as a predictor of non-Hodgkin's lymphoma
response to treatment with 3-(4-amino-l-oxo-l,3- dihydro-
isoindol-2-yl)-piperidine-2,6-dione are also disclosed.
~1 Citing
209. Gene network inference and biochemical assessment delineates GPCR pathways and CREB targets in
small intestinal neuroendocrine neoplasia
By Drozdov, Ignat; Svejda, Bernhard; Gustafsson, Bjorn I.; Mane, Shrikant; Pfragner, Roswitha; Kidd, Mark; Modlin,
Irvin M.
Small intestinal (SI) neuroendocrine tumors (NET) are increasing in incidence, however little is known about their biol.
High throughput techniques such as inference of gene regulatory networks from microarray expts. can objectively define
signaling machinery in this disease. Genome-wide co-expression anal. was used to infer gene relevance network in SI-
NETs. The network was confirmed to be non-random, scale-free, and highly modular. Functional anal. of gene co-
expression modules revealed processes including 'Nervous system development', 'Immune response', and 'Cell-cycle'.
Importantly, gene network topol. and differential expression anal. identified over-expression of the GPCR signaling
regulators, the cAMP synthetase, ADCY2, and the protein kinase A, PRKAR1A. Seven CREB response element (CRE)
transcripts assocd. with proliferation and secretion: BEX1, BICD1, CHGB, CPE, GABRB3, SCG2 and SCG3 as well as
ADCY2 and PRKAR1A were measured in an independent SI dataset (n = 10 NETs; n = 8 normal prepns.). All were up-
regulated (p<0.035) with the exception of SCG3 which was not differently expressed. Forskolin (a direct cAMP activator,
10-5 M) significantly stimulated transcription of pCREB and 3/7 CREB targets, isoproterenol (a selective β-adrenergic
receptor agonist and cAMP activator, 10-5 M) stimulated pCREB and 4/7 targets while BIM-53061 (a dopamine D2 and
Serotonin [5-HT2] receptor agonist, 10-6 M) stimulated 100% of targets as well as pCREB; CRE transcription correlated
with the levels of cAMP accumulation and PKA activity; BIM-53061 stimulated the highest levels of cAMP and PKA (2.8-
fold and 2.5-fold vs. 1.8-2-fold for isoproterenol and forskolin). Gene network inference and graph topol. anal. in SI NETs
suggests that SI NETs express neural GPCRs that activate different CRE targets assocd. with proliferation and secretion.
In vitro studies, in a model NET cell system, confirmed that transcriptional effects are signaled through the
cAMP/PKA/pCREB signaling pathway and that a SI NET cell line was most sensitive to a D2 and 5-HT2 receptor agonist
BIM-53061.
~12 Citings
210. Separation of gametes containing abnormal nucleic acids involved in a genetic disease or multifactorial
disorder from healthy gametes containing normal nucleic acids
By Vega, Manuel; Drittanti, Lila; Goossens, Michel
SciFinder® Page 85
A method of sepg. gametes of a subject comprises discriminating a first population of gametes contg. an abnormal
nucleic acid sequence involved in a genetic disease or in a multifactorial disorder in the offspring of the subject, from a
second population of gametes which does not contain said abnormal nucleic acid sequence. In general, the method
comprises a step contacting the gametes with at least one discriminating substance capable of (i) altering or destroying
the first population of gametes, or of (ii) allowing the identification of the first or the second population of gametes. Thus,
ejaculated sperm cells from a healthy carrier heterozygous for the ∆F508 mutation on the CFTR gene are centrifuged on
a d. gradient made of SupraSperm in order to obtain the high mobility fraction composed of viable spermatozoids, and
then labeled with an anti-CFTR monoclonal antibody (the discriminating substance) and stained to visualize the nuclei.
Green fluorescence reveals the presence of the CFTR protein, and blue fluorescence reveals the presence of the nuclei.
The sperm of the healthy carrier, heterozygous for the ∆508 mutation, contains both types of spermatozoids: those
expressing the CFTR protein (green and blue fluorescences) and those which do not express the CFTR protein (blue
fluorescence only). Sperm cells recovered from the high mobility fraction after centrifugation in a d. gradient were
incubated with a first anti-CFTR monoclonal antibody and with a second antibody directed against said first antibody.
Sperm cells were then sepd. by fluorescence activated cell sorting. The present invention offers a novel and unique
approach, in the context of preventive medicine, to avoid the transmission of a deleterious trait leading to a genetic or
multifactorial disease in the offspring of a subject, and limits or avoids preimplantation genetic diagnosis, invasive testing
during pregnancy, as well as pregnancy interruption or termination.
~0 Citings
211. The flavonone naringenin inhibits chloride secretion in isolated colonic epithelia
By Collins, Danielle; Kopic, Sascha; Geibel, John P.; Hogan, Aisling M.; Medani, Mekki; Baird, Alan W.; Winter,
Desmond C.
From European Journal of Pharmacology (2011), 668(1-2), 271-277. Language: English, Database: CAPLUS,
DOI:10.1016/j.ejphar.2011.06.052
Studies investigating the activating and inhibitory actions of bioflavonoids on colonic function have yielded conflicting
results. At low concns., flavonoids may stimulate Cl- secretion while at higher concns. they may have antisecretory
actions in the colon. Naringenin (4',5,7-trihydroxyflavanone), found predominantly in citrus fruits, confers a protective
effect against colorectal cancer and is purported to modulate secretory function in colonic cell lines. The aim of this study
was to investigate the effects of naringenin on ion transport in rat and human colonic mucosae. Naringenin inhibited
basal and stimulated chloride secretion in rat and human colonic mucosae mounted in Ussing chambers (IC50 330µMol/L
and 360µMol/L resp.) and did not alter intracellular cAMP generation. Naringenin inhibited Cl- secretion in MQAE (N-
(ethoxycarbonylmethyl)-6-methoxyquinolinium bromide) loaded crypts stimulated with forskolin. In BCECF (2',7'-bis-(2-
carboxyethyl)-5-(and 6)-carboxyfluorescein acetoxymethyl ester) loaded crypts, naringenin caused an intracellular
acidification (∆pH/min = 0.05 ± 0.004) which was sensitive to the Na-K-Cl co-transporter (NKCC) inhibitor bumetanide. In
addn., the antisecretory effect of naringenin was not inhibited by blockade of Ba sensitive basolateral K+ transporters or
by inhibition of Na+/H+ exchange by amiloride. We propose that the antisecretory action of naringenin is due to inhibition
of basolateral NKCC1 in rat and human colon.
~2 Citings
212. Treating cancer with statins and compounds having dipyridamole activity
By Penn, Linda; Schimmer, Aaron; Pandyra, Aleksandra
The disclosure pertains to methods of treating a cancer comprising administering to a subject in need thereof an effective
amt. of a statin in combination with an effective amt. of a dipyridamole and/or a compd. that has dipyridamole activity.
~1 Citing
213. Berberine reduces cAMP-induced chloride secretion in T84 human colonic carcinoma cells through
inhibition of basolateral KCNQ1 channels
By Alzamora, Rodrigo; O'Mahony, Fiona; Ko, Wing-Hung; Yip, Tiffany Wai-Nga; Carter, Derek; Irnaten, Mustapha;
Harvey, Brian Joseph
From Frontiers in Membrane Physiology and Biophysics (2011), 2(June), 33. Language: English, Database: CAPLUS,
DOI:10.3389/fphys.2011.00033
SciFinder® Page 86
Berberine is a plant alkaloid with multiple pharmacol. actions, including antidiarrheal activity and has been shown to
inhibit Cl- secretion in distal colon. The aims of this study were to det. the mol. signaling mechanisms of action of
berberine on Cl- secretion and the ion transporter targets. Monolayers of T84 human colonic carcinoma cells grown in
permeable supports were placed in Ussing chambers and short-circuit current measured in response to secretagogues
and berberine. Whole-cell current recording swere performed in T84 cells using the patch-clamp technique. Berberine
decreased forskolin-induced short-circuit current in a concn.-dependent manner (IC50 80 ± 8 µM). In apically
permeabilized monolayers and whole-cell current recordings, berberine inhibited a cAMP-dependent and chromanol
293B-sensitive basolateral membrane K+ current by 88%, suggesting inhibition of KCNQ1 K+ channels. Berberine did
not affect either apical Cl- conductance or basolateral Na+-K+-ATPase activity. Berberine stimulated p38 MAPK, PKCα
and PKA, but had no effect on p42/p44 MAPK and PKCδ. However, berberine pre-treatment prevented stimulation of
p42/p44 MAPK by epidermal growth factor. The inhibitory effect of berberine on Cl- secretion was partially blocked by
HBDDE (∼65%), an inhibitor of PKCα and to a smaller extent by inhibition of p38 MAPK with SB202190 (∼15%).
Berberine treatment induced an increase in assocn. between PKCα and PKA with KCNQ1 and produced
phosphorylation of the channel. We conclude that berberine exerts its inhibitory effect on colonic Cl- secretion through
inhibition of basolateral KCNQ1 channels responsible for K+ recycling via a PKCα-dependent pathway.
~2 Citings
214. PI3K modulators, Rho kinase modulators and methods of identifying and using same
By Fang, Ye; Sun, Haiyan; Deng, Huayun; Ferrie, Ann M.; Tran, Elizabeth
Disclosed are methods to characterize PI3K inhibitors and Rho kinase inhibitors using label-free cellular assays.
Disclosed are also methods to characterize a cell as to whether it has a deregulated PI3K pathway or not. To correctly
detect the phenotypic pharmacol. of a ROCK inhibitor that acts via the inhibition of ROCK activity in a live cell, one needs
a panel of cells as well as panels of markers using the biosensor cellular assays. Thus, a ROCK inhibitor responsive cell
line, A549, and a ROCK inhibitor unresponsive cell line, HepG2, were chosen as model systems. A panel of markers for
A549 cells was also chosen: pinacidil (an ATP-sensitive potassium ion channel opener), poly(I:C) (an agonist for tolyl-like
receptor subtype 3, TLR3), PMA (a small mol. activator for protein kinase C), SLIGKV-amide (an agonist for endogenous
protease activated receptor subtype 2), forskolin (an activator for adenylylcyclase), and histamine (an agonist for
endogenous histamine receptors). All of these markers led to dose-dependent and saturable biosensor signals, but with
distinct dynamics and characteristics (data not shown). The concns. closer to EC 100 were chosen for these markers for
generating the biosensor modulation indexes of any mol., particularly ROCK inhibitors. Using the panel of cells/markers,
over 500 compds., including ROCK inhibitor Y-27632, were systematically characterized.
~0 Citings
215. Active substance combination with gemcitabine for the treatment of epithelial cancer
By Heeschen, Christopher; Mueller, Maria Theresa; Hermann, Patrick Christian; Huber, Stephan
The present invention refers to active substance combinations
comprising of a nucleoside analog or antimetabolic agent like
Gemcitabine, and either a Nodal/Activin inhibitor or a SHH-
Inhibitor and an mTOR-inhibitor, medicaments comprising the
same and the use of the active substance combinations in the
treatment of cancer, esp. of epithelial cancer.
~1 Citing
216. Promoter-specific effects of metformin on aromatase transcript expression

SciFinder® Page 87
By Samarajeewa, Nirukshi U.; Ham, Seungmin; Yang, Fangyuan; Simpson, Evan R.; Brown, Kristy A.
From Steroids (2011), 76(8), 768-771. Language: English, Database: CAPLUS, DOI:10.1016/j.steroids.2011.02.041
Phase III aromatase inhibitors (AIs) are proving successful in the treatment of hormone-dependent postmenopausal
breast cancer. Side-effects assocd. with total body aromatase inhibition have prompted new research into the
development of breast-specific AIs. The identification of tissue- and disease-specific usage of aromatase promoters has
made the inhibition of aromatase at the transcriptional level an interesting approach. We have previously demonstrated
that AMPK-activating drugs, including metformin, were potent inhibitors of aromatase expression in primary human
breast adipose stromal cells (hASCs). This study examines the promoter-specific effects of metformin on inhibiting
aromatase expression in hASCs. Tumor-assocd. promoters PII/PI.3 were activated using forskolin (FSK)/phorbol ester
(PMA), whereas normal adipose assocd. promoter PI.4 was activated using dexamethasone (DEX)/tumor necrosis
factor-α (TNFα). Results demonstrate that metformin significantly decreased the FSK/PMA-, but not the DEX/TNFα-
mediated expression of total aromatase at concns. of 10, 20, and 50 µM. Using PCR to amplify promoter-specific
transcripts of aromatase, it appears that the inhibition of the FSK/PMA-mediated expression of aromatase is due to
decreases in PII/PI.3-specific transcripts, whereas no effect of metformin is obsd. on any promoter-specific transcript,
including PI.4, in DEX/TNFα-treated hASCs. This report therefore supports the hypothesis that metformin would act as a
breast-specific inhibitor of aromatase expression in the context of postmenopausal breast cancer.
~14 Citings
217. Tissue-specific regulation of aromatase promoter II by the orphan nuclear receptor LRH-1 in breast adipose
stromal fibroblasts
By Chand, Ashwini L.; Herridge, Kerrie A.; Howard, Tamara L.; Simpson, Evan R.; Clyne, Colin D.
From Steroids (2011), 76(8), 741-744. Language: English, Database: CAPLUS, DOI:10.1016/j.steroids.2011.02.024
In postmenopausal breast cancers, the increase in aromatase expression obsd. in tumor assocd. adipose stromal cells is
mediated via the upregulation of promoter II (PII) transcription. Factors such as PGE2 which are secreted from breast
carcinomas induce PII expression. The orphan nuclear receptor LRH-1/NR5A2 is one of the crit. downstream
transcriptional mediators of this effect. The aim of the current study was to det. whether LRH-1 could bind directly to PII
and whether the suppression of LRH-1 expression could inhibit aromatase expression in human adipose stromal
fibroblasts. Chromatin immunopptn. demonstrated endogenous LRH-1 occupancy on PII under basal conditions and
with treatment with forskolin and phorbol 12-myristate 13-acetate (PMA). To assess the impact of LRH-1 knockdown on
FSK/PMA mediated PII expression, cells were transfected with shRNA targeted against LRH-1 (shLRH-1) and treated
with forskolin and PMA. A decrease in LRH-1, PII and total aromatase mRNA transcripts was obsd. in shLRH-1
transfected cells compared to controls under basal and treatment conditions. The results of this study support the
hypothesis that suppression of LRH-1 may potentially be beneficial in the tissue specific regulation of aromatase
expression in post menopausal breast cancer.
~9 Citings
218. Cyclic AMP-induced p53 destabilization is independent of EPAC in pre-B acute lymphoblastic leukemia
cells in vitro
By Safa, Majid; Kazemi, Ahmad; Zaker, Farhad; Razmkhah, Farnaz
From Journal of Receptors and Signal Transduction (2011), 31(3), 256-263. Language: English, Database: CAPLUS,
DOI:10.3109/10799893.2011.578140
Context: Activation of the tumor suppressor protein p53 facilitates the cellular response to genotoxic stress. Thus,
releasing the wild-type p53 from indirect suppression would be crucial to successful killing of cancer cells by DNA-
damaging therapeutic agents. Objective: The aim of this study was to investigate the inhibitory role of cAMP levels on
p53 protein in acute lymphoblastic leukemia (ALL) cells. More importantly, we were interested to show through which
receptor cAMP acts to promote p53 degrdn. Materials and methods: In cell cultures, we investigated the effects of
forskolin/3-isobutyl-1-methylxanthine (IBMX) on stimulated p53 of ALL cell lines. Western blotting anal. was performed to
detect the expression of p53, phospho-p53, acetylated-p53, phospho-cAMP response element-binding protein (CREB),
and Mdm2 proteins. Flow cytometry was applied to analyze apoptosis. The gene expression of p53 and its target genes
was examd. by real-time polymerase chain reaction. Results: We show that elevation of cAMP levels in ALL cells
exposed to DNA damage attenuates p53 accumulation. Inhibition of proteasome function with MG-132 reversed the
inhibitory effect of cAMP on p53. However, targeting the p53--Mdm2 interaction did not rescue accumulated p53 from
the destabilizing signal of cAMP. The specific agonist of the cAMP receptor exchange protein activated by cAMP had no
effect on p53 expression in doxorubicin-treated NALM-6 cells, whereas PKA activators decreased p53 accumulation.
Discussion and conclusion: Our studies demonstrate that cAMP-PKA pathway regulates the sensitivity toward DNA-
damaging agents via inhibition of a p53-dependent pathway in B-cell precursor ALL (BCP-ALL) cells.
~4 Citings
SciFinder® Page 88
219. Prostaglandin E2 promotes the cell proliferation ability of CCLP1 through EP2 prostanoid receptor which
increased the expression of SnoN by activation of cAMP-PKA-CREB pathway
By Huang, Wenyong; Zhang, Li; Shu, Wei; Peng, Tao; Zhang, Hai; Ma, Juan; Bai, Xiaoming; Leng, Jing
From Nanjing Yike Daxue Xuebao (2011), 31(2), 143-148. Language: Chinese, Database: CAPLUS
The mechanism of prostaglandin E2 (PGE2) affecting the cell proliferation ability of human bile duct carcinoma cells
CCLP1 was explored. CCLP1 cell were treated with PGE2, EP1-4 receptor agonist, adenylate cyclase (AC) agonist
(Forskolin), PKA antagonist (H89), cAMP analogs (db-cAMP). The expression levels of ski-related novel gene (SnoN)
mRNA and SnoN protein, and the cell proliferation ability were examd. by RT-PCR, Western blot and WST methods in
CCLP1 cells, resp. The expression of SnoN mRNA in CCLP1 cells increased by 22.5% (P<0.01), while the level of SnoN
protein increased by 35.6% (P<0.05) after treated with 10 µmol/L PGE2 for 24 h. The expression of SnoN protein in
CCLP1 cells increased by 64.9% (P<0.05) after treated with EP2 receptor agonist (10 µmol/L) for 24 h, and cell
proliferation ability of CCLP1 increased by 26.5%. The levels of SnoN protein and CREB phosphorylation in CCLP1 cells
increased by 25.1% and 71.3% (P<0.05), resp., compared with the control group after treated with AC agonist Forskolin
(10 µmol/L) for 24 h, and the cell proliferation ability of CCLP1 increased by 4.4% (P<0.05) after treated with 10 µmol/L
Forskolin. The expression of SnoN in CCLP1 cells increased by 90.1% (P<0.05) when treated with cAMP analogs db-
cAMP (500 µmol/L) for 24 h, and decreased when treated with PKA antagonist H89 (10 µmol/L) for 24 h, the levels of
SnoN expression and CREB phosphorylation in CCLP1 cells treated with H89 decreased by 9.1% and 14.1% (P<0.05)
compared with those treated with Forskolin. Cell proliferation of CCLP1 decreased by 45.7% (P<0.05) after treated with
20 µmol/L H89. PGE2 might up-regulate the expression level of SnoN through EP2 receptor of CCLP1 cells which could
be partly related to the cAMP-PKA-CREB signaling pathway, and promote the cell proliferation.
~0 Citings
220. Rapid differentiation of human embryonal carcinoma stem cells (NT2) into neurons for neurite outgrowth
analysis
By Tegenge, Million Adane; Roloff, Frank; Bicker, Gerd
From Cellular and Molecular Neurobiology (2011), 31(4), 635-643. Language: English, Database: CAPLUS,
DOI:10.1007/s10571-011-9659-4
Human neurons derived from stem cells can be employed as in vitro models to predict the potential of neurochems.
affecting neurodevelopmental cellular processes including proliferation, migration, and differentiation. Here, we
developed a model of differentiating human neurons from well characterized human embryonal carcinoma stem cells
(NT2). NT2 cells were induced to differentiate into neuronal phenotypes after 2 wk of treatment with retinoic acid in
aggregate culture. Nestin pos. progenitor cells migrate out of NT2 aggregates and differentiate into βIII-tubulin
expressing neuronal cells. Culturing the NT2 cells for an addnl. 7-14 days resulted in increased percentage of βIII-tubulin
expressing cells, elaborating a long neurite that pos. stained for axonal marker (Tau) and presynaptic protein (synapsin).
We then asked whether neurite outgrowth from NT2 cells is modulated by bioactive chems. Since the cAMP/PKA
pathway has been widely investigated as a regulator of neurite outgrowth/regeneration in several exptl. systems, we
used chem. activators and inhibitors of cAMP/PKA pathway in the culture. The adenylyl cyclase activator, forskolin, and
cell-permeable analog of cAMP, 8-Br-cAMP increased the percentage of neurite bearing cells and neurite extension.
Application of the protein kinase A inhibitors, H-89 and Rp-cAMP, blocked neurite formation. Taken together, NT2
aggregates undergo migration, differentiation, and neurite elaboration and can be used as a model of differentiating
human neurons to screen neurochems. and to understand cellular mechanisms of human nerve cell development.
~13 Citings
221. Heterotrimeric stimulatory GTP-binding proteins inhibit cisplatin-induced apoptosis by increasing X-linked
inhibitor of apoptosis protein expression in cervical cancer cells
By Cho, Eun-Ah; Oh, Jung-Min; Kim, So-Young; Kim, Yeni; Juhnn, Yong-Sung
From Cancer Science (2011), 102(4), 837-844. Language: English, Database: CAPLUS, DOI:10.1111/j.1349-
7006.2011.01883.x
SciFinder® Page 89
Treatment with cisplatin (cis-dichlorodiammineplatinum (II)) induces DNA double-stranded breaks and apoptosis in many
human cancer cells. We have reported that heterotrimeric stimulatory GTP-binding proteins (Gαs) can modulate the
apoptotic response of several cancer cells. This study investigated the effect of Gαs on apoptosis triggered by cisplatin
and its underlying mol. mechanism in cervical cancer cells. Stable expression of constitutively active Gαs (GαsQL)
decreased the release of cytochrome c from the mitochondria to the cytosol and cleavage of caspase-3 and poly(ADP-
ribose) polymerases in HeLa cells treated with 30 µM cisplatin, indicating that Gαs inhibited cisplatin-induced apoptosis.
Treatment with forskolin also inhibited apoptosis of C33A and CaSKi cervical cancer cells. Expression of GαsQL
increased the expression of the X-linked inhibitor of apoptosis protein (XIAP) and partially maintained increased XIAP
after cisplatin treatment. Knockdown of XIAP by siRNA augmented apoptosis. Expression of GαsQL increased XIAP
mRNA; this increase was inhibited by a protein kinase A inhibitor and cAMP response element (CRE) decoy. A cAMP
response element (CRE)-like element at -1396 bp in the XIAP promoter was found to mediate the induction of XIAP by
Gαs. In addn., expression of GαsQL protected against the ubiquitin/proteasome-dependent degrdn. of the XIAP protein.
This study shows that Gαs inhibits cisplatin-induced apoptosis by increasing transcription of XIAP and by decreasing
degrdn. of XIAP protein in HeLa cervical cancer cells.
~9 Citings
222. Reprogramming of mouse fibroblasts to an intermediate state of differentiation by chemical induction

By Park, Joonghoon; Kim, Chul; Tang, Yong; Amano, Tomokazu; Lin, Chih-Jen; Tian, Cindy
From Cellular Reprogramming (2011), 13(2), 121-131. Language: English, Database: CAPLUS,
DOI:10.1089/cell.2010.0067
Induced pluripotent stem cells (iPSCs) generated by forced expression of four transcription factors offer promises for
regenerative and therapeutic uses in human diseases. However, it is necessary to overcome the risk of tumorigenicity
caused by the use of potent oncogenes and the use of randomly integrating vectors before the iPSC technol. can be
applied to human medicine. Stem cells and cancer cells share many features in common, implying that there are similar
underlying mechanisms in their development. Small mols. have been used to induce cell reprogramming for lineage
trans-differentiation and for maintaining pluripotency of stem cells. In this study, we investigated the possibility of
replacing all reprogramming viral factors with small mols. To this end, we evaluated the effects of carcinogens at
nongenotoxic levels on somatic cells. We identified 16 candidate chems. through biol.-oriented in silico high-throughput
screening with com. available inventories from Sigma-Aldrich for cancer research, and established a reprogramming
protocol of 16-day treatment followed by 5 days of recovery. This protocol was applied to B6/129 mouse embryonic
fibroblasts (MEFs) at passage 3. From recovery day 2, colonies appeared at an efficiency of 0.02%. These colonies
were pos. for both alk. phosphatase and surface specific embryonic antigen-1 (SSEA-1) at a comparable level to those of
mouse embryonic stem cells (ESCs). Global gene expression anal. with a 38K gene MEEBO microarray revealed that
the colonies had 564 (1.5%) differentially expressed genes compared to MEFs at day 0 of treatment, and these genes
were enriched in "neuromuscular differentiation.". Moreover, 122 differentially expressed genes in the colonies were
ESC-enriched, including downregulated somatic markers and upregulated stem cell markers. In conclusion, combined
chem. treatment of MEFs herein might have caused these cells to transverse to an intermediate state within the
mesodermal lineages.
~8 Citings
223. Role of histamine H4 receptor in breast cancer cell proliferation

By Medina, Vanina A.; Brenzoni, Pablo G.; Lamas, Diego J. Martinel; Massari, Noelia; Mondillo, Carolina; Nunez,
Mariel A.; Pignataro, Omar; Rivera, Elena S.
From Frontiers in Bioscience, Elite Edition (2011), E3(3), 1048-1060. Language: English, Database: CAPLUS
In order to better understand the role of histamine H4 (H4R) receptor in breast cancer, we studied the receptor expression
pattern, assocd. signal transduction pathway and biol. responses, in breast cancer cell lines with different malignant
characteristics. A different pattern of protein expression was obsd. in MDA-MB-231 compared to MCF-7 cells detd. by
western blot, exhibiting the presence of a diverse range of mol. wt. species of the H4R. H4R agonist reduced cyclic
adenosine monophosphate (cAMP) formation induced by forskolin only in MCF-7 cells. In MDA-MB-231 cells, H4R
agonists significantly decreased cell proliferation, augmented the Annexin-V and TdT-mediated UTP-biotin Nick End
labeling (TUNEL) pos. cells and produced a 2.5-fold increase in cell senescence. In MCF-7 cells, H4R agonists inhibited
proliferation by 50%, increasing the exponential doubling time. This effect was assocd. to an augment in Annexin-V and
TUNEL pos. cells, and a 2-fold increase in cell senescence. We conclude that H4R is functionally expressed in human
breast cancer cell lines, exhibiting a key role in histamine-mediated biol. processes such as cell proliferation, senescence
and apoptosis.
~0 Citings

SciFinder® Page 90
224. Norepinephrine induces VEGF expression and angiogenesis by a hypoxia-inducible factor-1α protein-
dependent mechanism
By Park, Soon-Young; Kang, Joo-Hee; Jeong, Kang-Jin; Lee, Jang-Soon; Han, Jeong-Whan; Choi, Wahn-Soo; Kim,
Yong-Kee; Kang, Jae-Ku; Park, Chang-Gyo; Lee, Hoi Young
From International Journal of Cancer (2011), 128(10), 2306-2316. Language: English, Database: CAPLUS,
DOI:10.1002/ijc.25589
A growing no. of studies have demonstrated that physiol. factors can influence the progression of several cancers via
cellular immune function, angiogenesis and metastasis. Recently, stress-induced catecholamines have been shown to
increase the expression of various cancer progressive factors, including vascular endothelial growth factor (VEGF),
matrix metalloproteinases and interleukins. However, a detailed mechanism remains to be identified. In this study, we
investigated the role of adrenergic receptors and hypoxia-inducible factor (HIF)-1α protein in catecholamine-induced
VEGF expression and angiogenesis. Treatment of the cells with norepinephrine (NE) or isoproterenol induced VEGF
expression and HIF-1α protein amt. in a dose-dependent manner. Induction of VEGF expression by NE was abrogated
when the cells were transfected with HIF-1α-specific siRNA. Similarly, adenylate cyclase activator forskolin and cAMP-
dependent protein kinase A inhibitor H-89 enhanced and decreased HIF-1α protein amt., resp. More importantly,
conditioned medium of NE-stimulated cancer cells induced angiogenesis in a HIF-1α protein-dependent manner. In
addn., pretreatment of cells with propranolol, a β-adrenergic receptor (AR) blocker, completely abolished induction of
VEGF expression and HIF-1α protein amt. by NE in all of the tested cancer cells. However, treatment with the α1-AR
blocker prazosin inhibited NE-induced HIF-1α protein amt. and angiogenesis in SK-Hep1 and PC-3 but not MDA-MB-231
cells. Collectively, our results suggest that ARs and HIF-1α protein have crit. roles in NE-induced VEGF expression in
cancer cells, leading to stimulation of angiogenesis. These findings will help to understand the mechanism of cancer
progression by stress-induced catecholamines and design therapeutic strategies for cancer angiogenesis.
~40 Citings
225. Substituted 3-aminocinnolin-4(1H)-one derivatives, their preparation, and their therapeutic application as
By Legeay, Carole; Rinaldi-Carmona, Murielle; Roux, Pascale; Vernhet, Claude
From PCT Int. Appl. (2011), WO 2011033225 A2 20110324, Language: French, Database: CAPLUS
The invention is related to the prepn. of title compds. I [R1 = H, alkyl; R2 = (un)substituted alkyl, non-arom. C3-12
carbocyclyl; or NR1R2 = (un)substituted azetidin-1-yl, pyrrolidin-1-yl, piperidin-1-yl, morpholin-4-yl, piperazin-1-yl; R3 =
(CH2)nR5; R4 = H, halo, Alk, OAlk; R5 = halo, CF3, S(O)0-2Alk, NR6SO2Alk; R6 = H, alkyl; Alk = C1-4 alkyl optionally
substituted with ≥1 F's; n = 2-5; and their acid addn. salts] as agonists of CB2 cannabinoid receptors and their use for
treatment of the diseases it implies (no data). Thus, a multi-step synthesis starting from 1-(2-aminophenyl)ethanone (II)
via cyclization of II in the presence of NaNO2 in concd. HCl for 2 h at 0° gave pentylcinnoline III (m.p. = 82-84°). I have
very good affinity in vitro for cannabinoid CB2 receptors (IC50< 500 nM). The agonist nature of compds. I was
demonstrated by adenylate-cyclase inhibition models (no data). The antagonist nature of compds. I was demonstrated in
models of reversion of the inhibition of adenylate cyclase (stimulated by forskolin) induced by an agonist of CB2
receptors (no data). The inverse agonist nature of compds. I was demonstrated in models of activation of adenylate
cyclase (stimulated by forskolin) (no data). I are useful for treating autoimmune, allergic, infection, neurodegenerative,
cardiovascular and gastrointestinal diseases, neoplasm, obesity and diabetes (no data).
SciFinder® Page 91
~0 Citings
226. Down-regulation of ICBP90 contributes to doxorubicin resistance

By Wang, Jingxuan; Song, Ying; Xu, Shanqi; Zhang, Qingyuan; Li, Yulian; Tang, Dabei; Jin, Shi
DOI:10.1016/j.ejphar.2011.01.042
Acquired resistance to doxorubicin has become a serious obstacle in breast cancer treatment. The underlying
mechanism responsible for this was not completely elucidated. In this study, a doxorubicin-resistant MCF-7/Dox cell was
developed to mimic the occurrence of acquired doxorubicin resistance. We next contrasted the expression profiles of
ICBP90 and Topo IIα and tumor cell growth of different breast cancer cell lines to doxorubicin. Decreased expression
levels of ICBP90 and Topo IIα were found in doxorubicin-resistant cells. To examine its function in chemoresistance,
RNA interference (RNAi) and forskolin stimulation expts. further demonstrated that ICBP90 and Topo IIα were involved in
the proliferation of cells that had acquired doxorubicin resistance. In MCF-7/Dox and ICBP90-siRNA cells, the cell
growth wasn't inhibited by doxorubicin and preferentially arrested in G1 phase. However, after forskolin increased the
Topo IIα expression, these breast cancer cells were again found to be inhibited by doxorubicin. Further,
immunohistochem. assay breast cancer patients accepted EFC regimen showed ICBP90 was significantly assocd. with
tumor cell proliferation, locally advanced disease, and Topo IIα expression. In conclusion, down-regulation of ICBP90
induced the descended expression of Topo IIα protein which is the target enzyme of doxorubicin.
~5 Citings
227. Study of 6-propyl-2-thiouracil as a radioprotector for the thyroid gland

SciFinder® Page 92
By Perona, Marina; Dagrosa, Alejandra; Pagotto, Romina; Casal, Mariana; Pignataro, Omar; Pisarev, Mario; Juvenal,
Guillermo
From International Journal of Low Radiation (2010), 7(5), 366-379. Language: English, Database: CAPLUS,
DOI:10.1504/IJLR.2010.036962
The objective of the paper was to study the application of 6-propyl-2-thiouracil (PTU) as a radioprotector for the thyroid
gland. Rat thyroid epithelial cells (FRTL-5) and human colon cancer cells (ARO81-1) were exposed to γ-irradn. with or
without 1 mM PTU. Radiation response was analyzed by clonogenic survival assay. CAMP levels were measured by
RIA. The results showed that PTU increased the Surviving Cell Fraction (SF) at 2 Gy significantly (p < 0.05) in both cell
lines. PTU increased extracellular levels of cAMP in all the treatments in a dose- and time-dependent manner for FRTL-
5 cells. In ARO81-1 cells, a peak was obsd. at 24 h in extracellular levels incubated with 1 mM PTU (36.97 ± 6.74
fmol/µg prot vs. control: 17.53 ± 3.9 fmol/µg prot, p < 0.001). Forskolin and dibutyril cAMP mimicked the effect of PTU on
SF. Thus PTU appears to be a radioprotector for thyroid cells and could exert its effect through cAMP.
~0 Citings
228. Substituted 3-aminocinnolin-4(1H)-one derivatives, their preparation, and their therapeutic application as
By Vernhet, Claude; Legeay, Carole; Roux, Pascale; Rinaldi Carmona, Murielle
The invention is related to the prepn. of title compds. I [R1 = H, alkyl; R2 = (un)substituted alkyl, non-arom. C3-12
carbocyclyl; or NR1R2 = (un)substituted azetidin-1-yl, pyrrolidin-1-yl, piperidin-1-yl, morpholin-4-yl, piperazin-1-yl; R3 =
(CH2)nR5, CH2R6; R4 = H, halo, Alk, OAlk; R5 = R4, S(O)0-2Alk, NR7SO2Alk; R6 = (un)substituted (hetero)aryl; R7 = H,
alkyl; Alk = C1-4 alkyl optionally substituted with ≥1 F's; n = 2-5; and their acid addn. salts] as agonists of CB2
cannabinoid receptors and their use for treatment of the diseases it implies (no data). Thus, a multi-step synthesis
starting from 1-(2-aminophenyl)ethanone (II) via cyclization of II in the presence of NaNO2 in concd. HCl for 2 h at 0°
gave pentylcinnoline III (m.p. = 82-84°). I have very good affinity in vitro for cannabinoid CB2 receptors (IC50< 500 nM).
The agonist nature of compds. I was demonstrated by adenylate-cyclase inhibition models (no data). The antagonist
nature of compds. I was demonstrated in models of reversion of the inhibition of adenylate cyclase (stimulated by
forskolin) induced by an agonist of CB2 receptors (no data). The inverse agonist nature of compds. I was demonstrated
in models of activation of adenylate cyclase (stimulated by forskolin) (no data). I are useful for treating autoimmune,
allergic, infection, neurodegenerative, cardiovascular and gastrointestinal diseases, neoplasm, obesity and diabetes (no
data).
SciFinder® Page 93
~0 Citings
229. A CRE that binds CREB and contributes to PKA-dependent regulation of the proximal promoter of human
RAB25 gene
By Xue, Huiping; Qiao, Yongxia; Ni, Peihua; Wang, Jiayi; Chen, Changqiang; Huang, Gang
RAB25 plays an important role in tumor progression and aggressiveness; altered RAB25 expression may cause human
cancer. As the underlying mechanism of RAB25-mediated carcinogenesis in various tumor types progressively comes to
light, RAB25 is expected to represent a novel therapeutic target. However, the regulation of RAB25 expression per se
has not yet been described. Here we have firstly identified and characterized the human RAB25 promoter. Using PCR-
based chromatin accessibility and chromatin immunopptn. (ChIP) assays, an open chromatin conformation (-173/+17)
was detected around the transcription start site of the RAB25 gene. Deletion constructs of the 5' flanking region were
fused to a luciferase reporter gene. After transient transfection in gastric cancer cell line AGS, a CRE (-67/-58) binding
CREB was identified in the core promoter region. Electrophoretic mobility shift (EMSA) and ChIP assays demonstrated
that CREB binds to the core promoter. Deletion of CREB consensus sequence resulted in the total loss of the promoter
activity. Moreover, we have also found forskolin, PKA activator, could enhance open chromatin accessibility, by which to
expose the CRE and facilitate phosphorylation of CREB, which in turn recruits co-factor CBP and Brg I and then results
in a more open chromatin configuration assocd. with local histone modification, finally heightening RAB25 expression
and strengthening its promoter activity. Therefore, the present study delineates the fundamental elements of a core
promoter structure that will be helpful for future studies regarding the regulation of RAB25 gene.
~8 Citings

SciFinder® Page 94
230. Signaling from the human melanocortin 1 receptor to ERK1 and ERK2 mitogen-activated protein kinases
involves transactivation of cKIT
By Herraiz, Cecilia; Journe, Fabrice; Abdel-Malek, Zalfa; Ghanem, Ghanem; Jimenez-Cervantes, Celia; Garcia-Borron,
Jose C.
From Molecular Endocrinology (2011), 25(1), 138-156. Language: English, Database: CAPLUS,
DOI:10.1210/me.2010-0217
Melanocortin 1 receptor (MC1R), a Gs protein-coupled receptor expressed in melanocytes, is a major determinant of skin
pigmentation, phototype and cancer risk. Upon stimulation by αMSH, MC1R triggers the cAMP and ERK1/ERK2 MAPK
pathways. In mouse melanocytes, ERK activation by αMSH binding to Mc1r depends on cAMP, and melanocytes are
considered a paradigm for cAMP-dependent ERK activation. However, human MC1R variants assocd. with red hair, fair
skin [red hair color (RHC) phenotype], and increased skin cancer risk display reduced cAMP signaling but activate ERKs
as efficiently as wild type in heterologous cells, suggesting independent signaling to ERKs and cAMP in human
melanocytes. We show that MC1R signaling activated the ERK pathway in normal human melanocytes and melanoma
cells expressing physiol. levels of endogenous RHC variants. ERK activation was comparable for wild-type and mutant
MC1R and was independent on cAMP because it was neither triggered by stimulation of cAMP synthesis with forskolin
nor blocked by the adenylyl cyclase inhibitor 2',5'-dideoxyadenosine. Stimulation of MC1R with αMSH did not lead to
protein kinase C activation and ERK activation was unaffected by protein kinase C inhibitors. Conversely, pharmacol.
interference, small interfering RNA studies, expression profiles, and functional reconstitution expts. showed that αMSH-
induced ERK activation resulted from Src tyrosine kinase-mediated transactivation of the stem cell factor receptor, a
receptor tyrosine kinase essential for proliferation, differentiation, and survival of melanocyte precursors, thus
demonstrating a functional link between the stem cell factor receptor and MC1R. Moreover, this transactivation
phenomenon is unique because it is unaffected by natural mutations impairing canonical MC1R signaling through the
cAMP pathway.
~35 Citings
231. Methods to characterize cell reprogramming using a panel of markers and generating primary profiles for
each marker
By Fang, Ye; Pai, Sadashiva K.; Verrier, Florence
Disclosed are label free biosensors and methods using these to observe stem cells and the for the anal. of stem and
related cells.
~1 Citing
232. Lubiprostone Activates Cl- Secretion via cAMP Signaling and Increases Membrane CFTR in the Human
Colon Carcinoma Cell Line, T84
By Ao, Mei; Venkatasubramanian, Jayashree; Boonkaewwan, Chaiwat; Ganesan, Nivetha; Syed, Asma; Benya,
Richard V.; Rao, Mrinalini C.
From Digestive Diseases and Sciences (2011), 56(2), 339-351. Language: English, Database: CAPLUS,
DOI:10.1007/s10620-010-1495-8
Background: Lubiprostone, used clin. (b.i.d.) to treat constipation, has been reported to increase transepithelial Cl-
transport in T84 cells by activating ClC-2 channels. Aim; To identify the underlying signaling pathway, we explored the
effects of short-term and overnight lubiprostone treatment on second messenger signaling and Cl- transport. Methods:
Cl- transport was assessed either as Isc across T84 monolayers grown on Transwells and mounted in Ussing chambers
or by the iodide efflux assay. [cAMP]i was measured by enzyme immunoassay, and [Ca2+]i by Fluo-3 fluorescence.
Quantitation of apical cell surface CFTR protein levels was assessed by Western blotting and biotinylation with the EZ-
Link Sulfo-NHS-LC-LC-Biotin. ClC-2 mRNA level was studied by RT-PCR. Results: Lubiprostone and the cAMP
stimulator, forskolin, caused comparable and maximal increases of Isc in T84 cells. The Isc effects of lubiprostone and
forskolin were each suppressed if the tissue had previously been treated with the other agent. These responses were
unaltered even if the monolayers were treated with lubiprostone overnight. Lubiprostone-induced increases in iodide
efflux were ∼ 80% of those obtained with forskolin. Lubiprostone increased [cAMP]i. H89, bumetanide, or CFTRinh-172
greatly attenuated lubiprostone-stimulated Cl- secretion, whereas the ClC-2 inhibitor CdCl2 did not. Compared to
controls, FSK-treatment increased membrane-assocd. CFTR by 1.9 fold, and lubiprostone caused a 2.6-fold increase in
apical membrane CFTR as seen by immunoblotting following cell surface biotinylation. Conclusions: Lubiprostone
activates Cl- secretion in T84 cells via cAMP, protein kinase A, and by increasing apical membrane CFTR protein.
~19 Citings

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233. Fibronectin increases RhoA activity through inhibition of PKA in the human gastric cancer cell line SGC-
7901
By Li, Yongjin; Chen, Yongchang; Tao, Yan; Wang, Ying; Chen, Yuefang; Xu, Wenrong
From Molecular Medicine Reports (2011), 4(1), 65-69. Language: English, Database: CAPLUS,
DOI:10.3892/mmr.2010.396
Fibronectin/integrin-mediated signaling plays a key role in the regulation of adhesion, migration and metastasis of
tumors. Numerous studies have addressed the significance of the assocn. between integrin and RhoA, but the exact
mechanism is unclear. Results from labs., including ours, have demonstrated that PKA inhibits the activity and function
of RhoA. This study was designed to investigate the relationships among the fibronectin/integrin-, cAMP/PKA- and
RhoA-mediated intracellular signal transduction pathways. Rho activity was detected by pull-down assay. CAMP concn.
was measured by RIA. The distribution of the PKA catalytic subunit and the phosphorylation of vasodilator-stimulated
phosphoprotein (VASP) were detected by fluorescence microscopy and Western blotting, resp., to examine the activation
of PKA. CAMP-mediated gene expression activity was analyzed using a luciferase reporter gene assay. The results
revealed that, in SGC-7901 cells, sol. fibronectin increased RhoA activity and blocked the inhibition of RhoA activity by
cAMP/PKA. The cAMP level, which was increased by forskolin and pertussis toxin, was decreased by fibronectin. The
nuclear location of the PKA catalytic unit, the phosphorylation of VASP and cAMP response element (CRE)-directed
reporter gene expression induced by forskolin were blocked by fibronectin. However, fibronectin did not block VASP
phosphorylation or CRE-directed reporter gene expression induced by cAMP. These data suggest that
fibronectin/integrin induces RhoA activation through the inhibition of cAMP/PKA signal transduction. The possible point
of action of fibronectin/integrin is adenylate cyclase.
~2 Citings
234. Frequent phosphodiesterase 11A gene (PDE11A) defects in patients with carney complex (CNC) caused by
PRKAR1A mutations: PDE11A may contribute to adrenal and testicular tumors in CNC as a modifier of the
phenotype
By Libe, Rossella; Horvath, Anelia; Vezzosi, Delphine; Fratticci, Amato; Coste, Joel; Perlemoine, Karine; Ragazzon,
Bruno; Guillaud-Bataille, Marine; Groussin, Lionel; Clauser, Eric; et al
CAPLUS, DOI:10.1210/jc.2010-1704
Background: Carney complex (CNC) is an autosomal dominant multiple neoplasia, caused mostly by inactivating
mutations of the regulatory subunit 1A of the protein kinase A (PRKAR1A). Primary pigmented nodular adrenocortical
disease (PPNAD) is the most frequent endocrine manifestation of CNC with a great inter-individual variability. Germline,
protein-truncating mutations of phosphodiesterase type 11A (PDE11A) have been described to predispose to a variety of
endocrine tumors, including adrenal and testicular tumors. Objectives: Our objective was to investigate the role of
PDE11A as a possible gene modifier of the phenotype in a series of 150 patients with CNC. Results: A higher frequency
of PDE11A variants in patients with CNC compared with healthy controls was found (25.3 vs. 6.8%, P < 0.0001). Among
CNC patients, those with PPNAD were significantly more frequently carriers of PDE11A variants compared with patients
without PPNAD (30.8 vs. 13%, P = 0.025). Furthermore, men with PPNAD were significantly more frequently carriers of
PDE11A sequence variants (40.7%) than women with PPNAD (27.3%) (P < 0.001). A higher frequency of PDE11A
sequence variants was also found in patients with large-cell calcifying Sertoli cell tumors (LCCSCT) compared with those
without LCCSCT (50 vs. 10%, P = 0.0056). PDE11A variants were significantly assocd. with the copresence of PPNAD
and LCCSCT in men: 81 vs. 20%, P < 0.004). The simultaneous inactivation of PRKAR1A and PDE11A by small
inhibitory RNA led to an increase in cAMP-regulatory element-mediated transcriptional activity under basal conditions
and after stimulation by forskolin. Conclusions: We demonstrate, in a large cohort of CNC patients, a high frequency of
PDE11A variants, suggesting that PDE11A is a genetic modifying factor for the development of testicular and adrenal
tumors in patients with germline PRKAR1A mutation.
~27 Citings
235. Tumor-secreted PGE2 Inhibits CCL5 Production in Activated Macrophages through cAMP/PKA Signaling
Pathway
By Qian, Xuesong; Zhang, Jidong; Liu, Jianguo
DOI:10.1074/jbc.M110.154971
SciFinder® Page 96
One of the major characteristics of tumors is their ability to evade immunosurveillance through altering the properties and
functions of host stromal and/or immune cells. CCL5 has been shown to play important roles in T cell proliferation, IFN-γ,
and IL-2 prodn., which promotes the differentiation and proliferation of Th1 cells important for immune defense against
intracellular infection. In this study we found that tumor-bearing mice were more susceptible to bacterial infection and
showed reduced CCL5 levels in serum during endotoxic shock. Our data further demonstrated that the sol. factors
secreted by mammary gland tumor cells but not normal mammary gland epithelial cells inhibited CCL5 expression in
macrophages in response to LPS, but not to TNF-α stimulation. The inhibitory effect of tumor-secreted mols. on LPS-
induced CCL5 expression was regulated at the post-transcriptional level. Blocking PGE2 synthesis by NS398 or through
the use of PGE2 receptor antagonists AH-6809 (EP2 antagonist) and AH-23848 (EP4 antagonist) completely reversed
the inhibitory effect of tumor-conditioned medium (TCM) on LPS-induced CCL5 expression. Moreover, PGE2 and the
cAMP analog forskolin could mimic tumor-mediated CCL5 inhibition, and the inhibitory effects of TCM, PGE2, and cAMP
analog on LPS-induced CCL5 expression could be completely reversed by the PKA inhibitor H89. Furthermore, blocking
PGE2 synthesis in vivo led to partial recovery of CCL5 prodn. during endotoxic shock. Taken together, our data indicate
that PGE2 secreted from breast cancer cells suppresses CCL5 secretion in LPS-activated macrophages through a
cAMP/PKA signaling pathway, which may result in suppression of host immune responses against subsequent bacterial
infection.
~29 Citings
236. The effect of valproate and levetiracetam on steroidogenesis in forskolin-stimulated H295R cells
By von Krogh, Kristine; Harjen, Hannah; Almaas, Camilla; Zimmer, Karin E.; Dahl, Ellen; Olsaker, Ingrid; Tauboll, Erik;
Ropstad, Erik; Verhaegen, Steven
From Epilepsia (2010), 51(11), 2280-2288. Language: English, Database: CAPLUS, DOI:10.1111/j.1528-
1167.2010.02702.x
Purpose: Endocrine disruptive effects have been frequently obsd. in patients using antiepileptic drugs (AEDs). Two
different AEDs, valproate (VPA) and levetiracetam (LEV), were tested in forskolin-stimulated human adrenal carcinoma
(H295R) cells to explore their effect on steroidogenesis. VPA has a long history as an anticonvulsant and is linked with
many of the endocrine disorders assocd. with AED use. LEV is a newer AED, and no endocrine disruptive effects have
been reported in humans to date. Methods: H295R cells, which are capable of full steroidogenesis, were stimulated with
forskolin and exposed to either VPA or LEV for 48 h. Medium was collected and analyzed for hormone prodn. For the
VPA-exposed cells, steroidogenic gene expression anal. was also conducted. Results: VPA exposure resulted in a
significant redn. in a significant redn. in progesterone and estradiol (E2) prodn., whereas testosterone (T) levels
remained unchanged. There were also significant alterations in expression level for most genes analyzed. LEV
exposure resulted in a minor, but statistically significant, redn. in T and E2 prodn. Discussion: Exposure of forskolin-
stimulated H295R cells to VPA led to an increased T/E2 ratio through a significant decrease in estradiol prodn. Gene
anal. suggested that VPA affects NR0BI expression. NR0BI inhibits promoters of other genes involved in
steroidogenesis, and the altered expression of NR0BI might explain the obsd. down-regulation in hormone prodn. The
effects of LEV exposure on hormone secretion were not considered to be biol. significant.
~4 Citings
237. cAMP and fibroblast growth factor 2 regulate bone sialoprotein gene expression in human prostate cancer
cells
By Li, Zhengyang; Sasaki, Yoko; Mezawa, Masaru; Wang, Shuang; Li, Xinyue; Yang, Li; Wang, Zhitao; Zhou, Liming;
Araki, Shouta; Matsumura, Hiroyoshi; et al
From Gene (2011), 471(1-2), 1-12. Language: English, Database: CAPLUS, DOI:10.1016/j.gene.2010.09.009
Bone sialoprotein (BSP) is a noncollagenous protein of the extracellular matrix in mineralized connective tissues that has
been implicated in the nucleation of hydroxyapatite. Forskolin (FSK), an activator of adenylate cyclase, increased the
intracellular cAMP level, which stimulates the proliferation and differentiation of osteoblasts. Fibroblast growth factor 2
(FGF2) is a potent mitogen in many cell types, including osteoblasts. In human prostate cancer DU145 cells, FSK (1 µM)
and FGF2 (10 ng/mL) increased BSP and Runx2 mRNA and protein levels at 3 and 12 h, resp. Transient transfection
analyses were performed using chimeric constructs of the human BSP gene promoter linked to a luciferase reporter
gene. Treatment of DU145 cells with FSK (1 µM) and FGF2 (10 ng/mL) increased the luciferase activities of constructs
between -60LUC to -927LUC and -108LUC to -927LUC, including the human BSP gene promoter. Effects of FSK and
FGF2 abrogated in constructs included 2 bp mutations in the two cAMP response elements (CRE1 and CRE2).
Luciferase activities induced by FSK and FGF2 were blocked by protein kinase A and tyrosine kinase inhibitors. Gel
mobility shift analyses showed that FSK and FGF2 increased the binding of CRE1 and CRE2. CRE1-protein complexes
were supershifted by phospho-CREB1 and c-Fos antibodies, and disrupted by CREB1, c-Jun, JunD, Fra2, p300, Runx2,
Dlx5 and Smad1 antibodies. CRE2-protein complexes were disrupted by CREB1, phospho-CREB1, c-Fos, c-Jun, JunD,
Fra2, p300, Runx2, Dlx5 and Smad1 antibodies. These studies demonstrate that FSK and FGF2 stimulate BSP
transcription in DU145 human prostate cancer cells by targeting the CRE1 and CRE2 elements in the human BSP gene
promoter.
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~7 Citings
238. Multiple Roles of Protein Kinase A in Arachidonic Acid-Mediated Ca2+ Entry and Tumor-Derived Human
Endothelial Cell Migration
By Fiorio Pla, Alessandra; Genova, Tullio; Pupo, Emanuela; Tomatis, Cristiana; Genazzani, Armando; Zaninetti,
Roberta; Munaron, Luca
From Molecular Cancer Research (2010), 8(11), 1466-1476. Language: English, Database: CAPLUS,
DOI:10.1158/1541-7786.MCR-10-0002
We recently showed that arachidonic acid (AA) triggers calcium signals in endothelial cells derived from human breast
carcinoma (B-TEC). In particular, AA-dependent Ca2+ entry is involved in the early steps of tumor angiogenesis in vitro.
Here, we investigated the multiple roles of the nitric oxide (NO) and cAMP/protein kinase A (PKA) pathways in AA-
mediated Ca2+ signaling in the same cells. B-TEC stimulation with 5 µmol/L AA resulted in endothelial NO synthase
(NOS) phosphorylation at Ser1177, and NO release was measured with the fluorescent NO-sensitive probe DAR4M-AM.
PKA inhibition by the use of the membrane-permeable PKA inhibitory peptide myristoylated PKI14-22 completely
prevented both AA- and NO-induced calcium entry and abolished B-TEC migration promoted by AA. AA-dependent
calcium entry and cell migration were significantly affected by both the NOS inhibitor NG-nitro-L-arginine Me ester and
the NO scavenger 2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl 3-oxide, suggesting that NO release is functionally
involved in the signaling dependent on AA. Moreover, pretreatment with carboxyamidotriazole, an antiangiogenic
compd. that interferes with agonist-activated calcium entry, prevented AA-dependent B-TEC motility. Interestingly, even
in the absence of AA, enhancement of the cAMP/PKA pathway with the adenylyl cyclase activator forskolin evoked a
calcium entry dependent on NOS recruitment and NO release. The functional relevance of AA-induced calcium entry
could be restricted to tumor-derived endothelial cells (EC) because AA evoked a smaller calcium entry in normal human
microvascular ECs compared with B-TECs, and even more importantly, it was unable to promote cell motility in wound
healing assay. This evidence opens an intriguing opportunity for differential pharmacol. treatment between normal and
tumor-derived human ECs.
~8 Citings
239. Evaluation of a bioluminescent mouse model expressing aromatase PII-promoter-controlled luciferase as a

tool for the study of endocrine disrupting chemicals
By Rivest, Patricia; Devine, Patrick J.; Sanderson, J. Thomas
From Toxicology and Applied Pharmacology (2010), 249(1), 33-40. Language: English, Database: CAPLUS,
DOI:10.1016/j.taap.2010.08.011
Dysfunction of the enzyme aromatase (CYP19) is assocd. with endocrine pathologies such as osteoporosis, impaired
fertility and development of hormone-dependent cancers. Certain endocrine disrupting chems. affect aromatase
expression and activity in vitro, but little is known about their ability to do so in vivo. We evaluated a bioluminescent
mouse model (LPTA)CD-1-Tg(Cyp19-luc)-Xen) expressing luciferase under control of the gonadal aromatase pII
promoter as an in vivo screening tool for chems. that may affect aromatase expression. We studied the effects of
forskolin, pregnant mare serum gonadotropin and atrazine in this model (atrazine was previously shown to induced pII-
promoter-driven aromatase expression in H295R human adrenocortical carcinoma cells). About 2-4 out of every group
of 10 male or female Cyp19-luc mice injected i.p. with 10 mg/kg forskolin had increased gonadal bioluminescence after
3-5 days compared to controls; the others appeared non-responsive. Similarly, about 4 per group of 9 individual females
injected with pregnant mare serum gonadotropin had increased ovarian bioluminescence after 24 h. There was a
statistically significant correlation between ovarian bioluminescence and plasma estradiol concns. (n = 14; p = 0.022).
Males exposed to a single dose of 100 mg/kg or males and females exposed to 5 daily injections of 30 mg/kg atrazine
showed no change in gonadal bioluminescence over a 7 day period, but a significant interaction was found between
atrazine (100 mg/kg) and time in female mice (p < 0.05; two-way ANOVA). Ex vivo luciferase activity in dissected organs
was increased by forskolin in testis, epididymis and ovaries. Atrazine (30 mg/kg/day) increased (30%) luciferase activity
significantly in epididymis only. In conclusion, certain individual Cyp19-luc mice are highly responsive to aromatase
inducers, suggesting this model, with further optimization, may have potential as an in vivo screening tool for
environmental contaminants.
~5 Citings
240. Substituted 1-alkylcinnolin-4(1H)-one derivatives, their preparation, and their therapeutic application as
SciFinder® Page 98
By Barbagallo, Elodie; Legeay, Carole; Rinaldi-Carmona, Murielle; Roux, Pascale; Vernhet, Claude
From PCT Int. Appl. (2010), WO 2010116084 A1 20101014, Language: French, Database: CAPLUS
The invention is related to the prepn. of title compds. I [X = (C2-5)alkylene optionally substituted with ≥1 Alk groups; Alk =
(C1-4)alkyl optionally substituted with ≥1 F atoms; R1 = (un)substituted Ph, naphthyl, pyridinyl, 1-benzothienyl, 1,3-
benzodioxolyl; R2 = H, halo, (un)substituted alkyl, alkylsulfonyl, alkylsulfanyl, etc.; R3 = H, halo, Alk, OAlk; and their acid
addn. salts] as agonists of CB2 cannabinoid receptors and their use for treatment of the diseases it implies (no data).
Thus, a multi-step synthesis starting from 2-amino-4-chlorobenzoic acid via cyclization of 1-(2-amino-4-
chlorophenyl)ethanone in the presence of NaNO2 in concd. HCl for 2 h at 0° gave pentylcinnoline II (m.p. = 82-84°). I
have very good affinity in vitro for cannabinoid CB2 receptors (IC50< 500 nM). The agonist nature of compds. I was
demonstrated by adenylate-cyclase inhibition models (no data). The antagonist nature of compds. I was demonstrated in
models of reversion of the inhibition of adenylate cyclase (stimulated by forskolin) induced by an agonist of CB2
receptors (no data). The inverse agonist nature of compds. I was demonstrated in models of activation of adenylate
cyclase (stimulated by forskolin) (no data). I are useful for treating autoimmune, allergic, infection, neurodegenerative,
cardiovascular and gastrointestinal diseases, neoplasm, obesity and diabetes (no data).
~0 Citings
241. Substituted 1-alkylcinnolin-4(1H)-one derivatives, their preparation, and their therapeutic application as
By Barbagallo, Elodie; Legeay, Carole; Rinaldi Carmona, Murielle; Roux, Pascale; Vernhet, Claude
The invention is related to the prepn. of title compds. I [X = (CH2)n; n = 2-5; R1 = (un)substituted Ph, naphthyl, pyridinyl,
1-benzothienyl, 1,3-benzodioxolyl; R2 = H, halo, (un)substituted alkyl, alkylsulfonyl, alkylsulfanyl, etc.; R3 = H, halo,
(un)substituted alkyl, alkoxy; and their acid addn. salts] as agonists of CB2 cannabinoid receptors and their use for
treatment of the diseases it implies (no data). Thus, a multi-step synthesis starting from 2-amino-4-chlorobenzoic acid
via cyclization of 1-(2-amino-4-chlorophenyl)ethanone in the presence of NaNO2 in concd. HCl for 2 h at 0° gave
pentylcinnoline II (m.p. = 82-84°). I have very good affinity in vitro for cannabinoid CB2 receptors (IC50< 500 nM). The
agonist nature of compds. I was demonstrated by adenylate-cyclase inhibition models (no data). The antagonist nature
of compds. I was demonstrated in models of reversion of the inhibition of adenylate cyclase (stimulated by forskolin)
induced by an agonist of CB2 receptors (no data). The inverse agonist nature of compds. I was demonstrated in models
of activation of adenylate cyclase (stimulated by forskolin) (no data). I are useful for treating autoimmune, allergic,
infection, neurodegenerative, cardiovascular and gastrointestinal diseases, neoplasm, obesity and diabetes (no data).
~0 Citings
SciFinder® Page 99
242. Expression of mTOR and downstream signalling components in the JEG-3 and BeWo human placental
choriocarcinoma cell lines
By Mparmpakas, Dionisis; Zachariades, Elena; Foster, Helen; Kara, Anjeli; Harvey, Amanda; Goumenou, Anastasia;
Karteris, Emmanouil
From International Journal of Molecular Medicine (2010), 25(1), 65-69. Language: English, Database: CAPLUS,
DOI:10.3892/ijmm_00000314
Emerging data suggest that nutritional status and body wt. are related to reproductive function, and nutrient imbalances
during pregnancy lead to changes in the expression of fetal genes. Recent studies show that the mTOR acts as a
placental growth signalling sensor and its expression is down-regulated in intrauterine growth restriction. To date, very
little is known about the expression of this signalling pathway in choriocarcinoma, one of the most lethal germ cell
cancers. In this study, cultures of fusigenic (BeWo) and non-fusigenic (JEG-3) human choriocarcinoma cell lines were
used to investigate the expression of mTOR and its downstream signalling components. The effects of an inducer of
syncytialisation (forskolin) on mTOR, eIF4E binding proteins (4EBPs) and ribosomal protein S6 kinases (S6Ks) in BeWo
cells were also assessed. RT-PCR studies revealed that mTOR, 4EBP and S6Ks are expressed at mRNA level in both
JEG-3 and BeWo cells. Semi-quant. RT-PCR anal. revealed that in early stages of syncytialisation (50 µM forskolin for
48 h), the expression of mTOR and 4EBP was down-regulated when compared to unstimulated cells. In fully
syncytialised cells (50 µM forskolin for 72 h) the expression of both genes was similar to basal levels. Interestingly, the
phosphorylation (Ser371, Thr389) status of p70S6K remained unaltered upon forskolin treatment. These data validate
BeWo cells as an exptl. model to study the effects of forskolin-induced syncytialisation on mTOR signalling.
~5 Citings
243. Leptin potentiates antiproliferative action of cAMP elevation via protein kinase A down-regulation in breast
cancer cells
By Naviglio, Silvio; Di Gesto, Davide; Illiano, Fausto; Chiosi, Emilio; Giordano, Antonio; Illiano, Gennaro; Spina,
Annamaria
DOI:10.1002/jcp.22288
Previously, we have shown that leptin potentiates the antiproliferative action of cAMP elevating agents in breast cancer
cells and that the protein kinase A (PKA) inhibitor KT-5720 prevented the antiproliferative effects induced by the leptin
plus cAMP elevation. The present expts. were designed to gain a better understanding about the PKA role in the
antitumor interaction between leptin and cAMP elevating agents and on the underlying signaling pathways. Here we
show that exposure of MDA-MB-231 breast cancer cells to leptin resulted in a strong phosphorylation of both ERK1/2
and STAT3. Interestingly, intracellular cAMP elevation upon forskolin pretreatment completely abrogated both ERK1/2
and STAT3 phosphorylation in response to leptin and was accompanied by a consistent CREB phosphorylation.
Notably, leptin plus forskolin cotreatments resulted in a strong decrease of both PKA regulatory RIα and catalytic
subunits protein levels. Importantly, pretreatment with the PKA inhibitor KT-5720 blocked the forskolin-induced CREB
phosphorylation and prevented both the inhibition by forskolin of leptin-induced ERK1/2 and STAT3 phosphorylation and
the PKA subunits down-regulation induced by the combination of leptin and forskolin. Altogether, our results indicate that
leptin-dependent signaling pathways are influenced by cAMP elevation and identify PKA as relevantly involved in the
pharmacol. antitumor interaction between leptin and cAMP elevating drugs in MDA-MB-231 cells. We propose a mol.
model by which PKA confers its effects. Potential therapeutic applications by our data will be discussed. J. Cell. Physiol.
225: 801-809, 2010. © 2010 Wiley-Liss, Inc.
~18 Citings
244. Cyclin D1 and cyclin D3 show divergent responses to distinct mitogenic stimulation
By Anderson, Alexandra A.; Child, Emma S.; Prasad, Aarathi; Elphick, Lucy M.; Mann, David J.
DOI:10.1002/jcp.22207
SciFinder® Page 100
D-type cyclins predominantly regulate progression through the cell cycle by their interactions with cyclin-dependent
kinases (cdks). Here, we show that stimulating mitogenesis of Swiss 3T3 cells with phorbol esters or forskolin can
induce divergent responses in the expression levels, localization and activation state of cyclin D1 and cyclin D3. Phorbol
ester-mediated protein kinase C stimulation induces S phase entry which is dependent on MAPK activation and
increases the levels and activation of cyclin D1, whereas forskolin-mediated cAMP-dependent protein kinase A
stimulation induces mitogenesis that is independent of MAPK, but dependent upon mTor and specifically increases the
level and activation of cyclin D3. These findings uncover addnl. levels of complexity in the regulation of the cell cycle at
the level of the D-type cyclins and thus may have important therapeutic implications in cancers where specific D-cyclins
are overexpressed. J. Cell. Physiol. 225: 638-645, 2010. © 2010 Wiley-Liss, Inc.
~2 Citings
245. Use of hypoxanthine for promotion of neuronal outgrowth

By Irwin, Carleen Ann
Disclosed herein is a method of promoting neuronal outgrowth in a neuron. The method comprises contacting the
neuron with an effective amt. of hypoxanthine, to thereby promote neuronal outgrowth of the neuron. The hypoxanthine
may be contacted in the absence of xanthine oxidase and/or in the absence of exogenous nerve growth factor (NGF),
and/or in the absence of exogenous D-mannose, and/or in the absence of exogenous oncomodulin, and/or in the
absence of exogenous TGF-B. The neuron may be an optic nerve neuron or a retinal neuron and/or an injured neuron.
Neurons may be from the central nervous system (CNS) or the peripheral nervous system (PNS). The methods are
useful for treating an injured neuron, for example to an optic nerve neuron, resulting from branch and central vein/artery
occlusion, trauma, edema, angle-closure glaucoma, open-angle glaucoma. Methods of treatment of various neurol.
injuries and diseases, as well as therapeutic compns., are also disclosed.
~0 Citings
246. Sunitinib Inhibits MEK/ERK and SAPK/JNK Pathways and Increases Sodium/Iodide Symporter Expression
in Papillary Thyroid Cancer
By Fenton, Mike S.; Marion, Kenneth M.; Salem, Andrew K.; Hogen, Rachel; Naeim, Faramarz; Hershman, Jerome M.
From Thyroid (2010), 20(9), 965-974. Language: English, Database: CAPLUS, DOI:10.1089/thy.2010.0008
Background: Sunitinib malate (Sutent, Pfizer, Inc.; SU11248) is a selective, multitargeted inhibitor of receptor tyrosine
kinases and has been shown to inhibit receptors for VEGF, PDGF, KIT, FLT3, and RET. The objective of this study was
to det. the effects of sunitinib on signal transduction pathways and on gene expression of iodide-metabolizing proteins in
papillary cancer cells with the RET/PTC1 rearrangement. Methods: We investigated the effects of sunitinib on cell
growth, signal transduction pathways, and thyroid-specific gene expression in papillary thyroid cancer (PTC) cell lines
that had the RET/PTC1 rearrangement. Results: Sunitinib inhibited proliferation of RET/PTC1 subclones in a time- and
dose-related manner. The mean 50% lethal concn. in the RET/PTC1 subclones was 1.81 µM. Incubation of RET/PTC1
cells with 1 µM sunitinib inhibited their migration potential and transformed their morphol. Sunitinib inhibited RET
autophosphorylation at Y1062 and the activation of signal transducer and activator of transcription 3 by blocking Y705
phosphorylation. Sunitinib caused cell cycle arrest in the G0/G1 phase and dephosphorylation of retinoblastoma protein,
but did not induce apoptosis. Western blot anal. of the p38, MEK/ERK, and SAPK/JNK mitogen-activated protein kinase
signal transduction pathways showed that sunitinib blocked ERK 1/2 and JNK phosphorylation in the cytoplasm.
Sunitinib treatment of RET/PTC1 cell lines, in combination, with forskolin induced expression of the sodium (Na)/iodide (I)
symporter (NIS) and the transcription factors that bind the NIS upstream enhancer. Mechanistically, the inhibition of both
MEK/ERK and SAPK/JNK cytoplasmic pathways individually and in combination caused an increase in NIS gene
expression. Conclusion: Sunitinib appears to target the cytosolic MEK/ERK and SAPK/JNK pathways in the RET/PTC1
cell lines, suggesting that blocking these pathways is at least part of the mechanism by which sunitinib inhibits cell
proliferation and causes stimulation of NIS gene expression in RET/PTC1 cells.
~23 Citings
247. Rhinovirus-induced exacerbations of asthma: how is the β2-adrenoceptor implicated?

By Trian, Thomas; Moir, Lyn M.; Ge, Qi; Burgess, Janette K.; Kuo, Curtis; King, Nicholas J. C.; Reddel, Helen K.;
Black, Judith L.; Oliver, Brian G.; McParland, Brent E.
From American Journal of Respiratory Cell and Molecular Biology (2010), 43(2), 227-233. Language: English,
Database: CAPLUS, DOI:10.1165/rcmb.2009-0126OC
Rhinovirus (RV) infections are the major cause of asthma exacerbations in children and adults. Under normal
circumstances, asthmatic airway obstruction improves spontaneously or characteristically briskly in response to inhaled
β2-adrenergic receptor (β2AR) agonists. During virus-assocd. exacerbations, an impaired response to β2AR agonists is
obsd.; the reason for this is not known. The objective of this study was to det. the effect of RV infection on airway
smooth muscle β2AR function. The human cell line Beas-2B and primary human bronchial epithelial cells (HBECs) were
infected with RV (multiplicity of infection = 1). After 1 or 5 days for primary and Beas-2B cells, resp., cell culture
supernatants were harvested, UV-irradiated to inactivate RV, and applied to human airway smooth muscle cells for 3
days to assess modifications of β2AR function. RV conditioned medium from Beas-2B and HBECs decreased β2AR
agonist-induced cAMP by 50 and 65%, resp. (n = 5; P < 0.05). When cAMP was induced independently of the β2AR
using forskolin, no impairment was found. Using flow cytometry, we demonstrated that this decrease was likely the result
of β2AR desensitization because membrane but not total cell receptor β2AR was decreased. Pretreatment of HBECs and
Beas-2B cells but not human airway smooth muscle cells with the corticosteroids dexamethasone or fluticasone
abolished virus-mediated β2AR loss of function. This study shows that epithelial infection with RV induces a decrease of
β2AR function on airway smooth muscle cells, potentially explaining the clin. observation of loss of β2AR agonist function
during RV-induced asthma exacerbations.
~9 Citings
248. Defective cAMP generation underlies the sensitivity of CNS neurons to neurofibromatosis-1 heterozygosity
By Brown, Jacquelyn A.; Gianino, Scott M.; Gutmann, David H.
From Journal of Neuroscience (2010), 30(16), 5579-5589. Language: English, Database: CAPLUS,
DOI:10.1523/JNEUROSCI.3994-09.2010
Individuals with the neurofibromatosis type 1 (NF1) inherited cancer syndrome exhibit neuronal dysfunction that
predominantly affects the CNS. In this report, we demonstrate a unique vulnerability of CNS neurons, but not peripheral
nervous system (PNS) neurons, to reduced Nf1 gene expression. Unlike dorsal root ganglion neurons, Nf1 heterozygous
(Nf1+/-) hippocampal and retinal ganglion cell (RGC) neurons have decreased growth cone areas and neurite lengths,
and increased apoptosis compared to their wild-type counterparts. These abnormal Nf1+/- CNS neuronal phenotypes do
not reflect Ras pathway hyperactivation, but rather result from impaired neurofibromin-mediated cAMP generation. In
this regard, elevating cAMP levels with forskolin or rolipram treatment, but not MEK (MAP kinase kinase) or PI3-K
(phosphatidylinositol 3-kinase) inhibition, reverses these abnormalities to wild-type levels in vitro. In addn., Nf1+/- CNS,
but not PNS, neurons exhibit increased apoptosis in response to excitotoxic or oxidative stress in vitro. Since children
with NF1-assocd. optic gliomas often develop visual loss and Nf1 genetically engineered mice with optic glioma exhibit
RGC neuronal apoptosis in vivo, we further demonstrate that RGC apoptosis resulting from optic glioma in Nf1
genetically engineered mice is attenuated by rolipram treatment in vivo. Similar to optic glioma-induced RGC apoptosis,
the increased RGC neuronal death in Nf1+/- mice after optic nerve crush injury is also attenuated by rolipram treatment
in vivo. Together, these findings establish a distinctive role for neurofibromin in CNS neurons with respect to vulnerability
to injury, define a CNS-specific neurofibromin intracellular signaling pathway responsible for neuronal survival, and lay
the foundation for future neuroprotective glioma treatment approaches.
~30 Citings
249. Metformin inhibits aromatase expression in human breast adipose stromal cells via stimulation of AMP-
activated protein kinase
By Brown, Kristy A.; Hunger, Nicole I.; Docanto, Maria; Simpson, Evan R.
DOI:10.1007/s10549-010-0834-y
AMP-activated protein kinase (AMPK) is recognized as a master regulator of energy homeostasis. In concert with the
AMPK-kinase LKB1, it has been shown to provide a mol. link between obesity and postmenopausal breast cancer via its
actions to inhibit aromatase expression, hence estrogen prodn., within the breast. The anti-diabetic drug metformin is
known to increase the activity of AMPK and was therefore hypothesized to inhibit aromatase expression in primary
human breast adipose stromal cells. Results demonstrate that metformin significantly decreases the forskolin/phorbol
ester (FSK/PMA)-induced expression of aromatase at concns. of 10 and 50 µM. Consistent with the hypothesized
actions of metformin to increase AMPK activity, treatment with 50 µM metformin results in a significant increase in
phosphorylation of AMPK at Thr172. Interestingly, metformin also causes a significant increase in LKB1 protein
expression and promoter activity, thereby providing for the first time an addnl. mechanism by which metformin activates
AMPK. Furthermore, metformin inhibits the nuclear translocation of CRTC2, a CREB-coactivator known to increase
aromatase expression which is also a direct downstream target of AMPK. Overall, these results suggest that metformin
would reduce the local prodn. of estrogens within the breast thereby providing a new key therapeutic tool that could be
used in the neoadjuvant and adjuvant settings and conceivably also as a preventative measure in obese women.
~59 Citings
250. Methods to accelerate the isolation of novel cell lines with limited potential for differentiation from
pluripotent stem cells
By West, Michael D.; Sargent, Geoffrey; Murai, James T.; Kessler, Steven; Chapman, Karen; Larocca, David
Methods of inducing the differentiation of pluripotent primordial stem cells, such as human embryonic stem cells, human
embryonic germ cells, human embryo-derived cells and human embryonal carcinoma cells are described. Methods of
identifying and isolating these subpopulations from mixed cultures are described. These subpopulations of cells possess
reduced differentiation potential compared to the original pluripotent stem cells and each subpopulation is capable of
being propagated 20 or more population doublings.
~3 Citings
251. Discovery of selective glucocorticoid receptor modulators by multiplexed reporter screening and their use
in treatment of inflammatory disorders
By Diamond, Marc; Gerber, Anthony
The invention relates to assays to detect selective gene regulation by ligand dependent transcription factors. The
invention also relates to selective modulators of the glucocorticoid receptor for treatment of inflammation and allergic and
immune-mediated diseases. Glucocorticoids are widely used to suppress inflammation and treat various allergic and
immune-mediated diseases. Some glucocorticoid receptor (GR) regulated genes mediate the therapeutic response,
while others cause debilitating side effects. To discover selective modulators of GR, we developed a high throughput,
multiplexed system to monitor GR regulation of four promoters simultaneously. An initial screen of 1040 natural products
and FDA-approved compds. identified selective modulators of GR signaling. Our results have important implications for
developing novel GR modulators and identifying crosstalk pathways that control selective GR gene regulation.
~0 Citings
252. Proteomic analysis reveals ATP-dependent steps and chaperones involvement in luteolin-induced lung
cancer CH27 cell apoptosis
By Lee, Hong-Zin; Yang, Wen-Hui; Bao, Bo-Ying; Lo, Pei-Ling
DOI:10.1016/j.ejphar.2010.05.053
The present study applied 2D electrophoresis to analyze the proteins involved in luteolin (50 µM)-induced CH27 cell
apoptosis. We found 7 proteins to be markedly changed. According to the data of anal. of these protein spots, we
hypothesized that ATP synthetic pathway and heat shock proteins were involved in luteolin-induced CH27 cell apoptosis.
In this study, luteolin induced a significant change in intracellular ATP levels and mitochondrial activity of CH27 cells.
Further expts. demonstrated that pretreatment with forskolin blocked the luteolin-induced cell death. P38 and heat shock
protein 27 may be important participants in the luteolin-induced changes in organization of actin microfilaments in this
study. In addn., endoplasmic reticulum stress is also important in the luteolin-induced CH27 cell apoptosis. Our findings
suggested that the function of mitochondria and endoplasmic reticulum is the integral factor in luteolin-induced CH27 cell
apoptosis.
~9 Citings
253. Estrogen and non-genomic upregulation of voltage-gated Na+ channel activity in MDA-MB-231 human
breast cancer cells: Role in adhesion
By Fraser, Scott P.; Ozerlat-Gunduz, Iley; Onkal, Rustem; Diss, James K. J.; Latchman, David S.; Djamgoz, Mustafa
B. A.
DOI:10.1002/jcp.22154
External (but not internal) application of β-estradiol (E2) increased the current amplitude of voltage-gated
Na+ channels
(VGSCs) in MDA-MB-231 human breast cancer (BCa) cells. The G-protein activator GTP-γ-S, by itself, also increased
the VGSC current while the G-protein inhibitor GDP-β-S decreased the effect of E2. Expression of GPR30 (a G-protein-
coupled estrogen receptor) in MDA-MB-231 cells was confirmed by PCR, Western blot and immunocytochem.
Importantly, G-1, a specific agonist for GPR30, also increased the VGSC current amplitude in a dose-dependent manner.
Transfection and siRNA-silencing of GPR30 expression resulted in corresponding changes in GPR30 protein expression
but only internally, and the response to E2 was not affected. The protein kinase A inhibitor, PKI, abolished the effect of
E2, while forskolin, an adenylate cyclase activator, by itself, increased VGSC activity. On the other hand, pre-incubation
of the MDA-MB-231 cells with brefeldin A (a trans-Golgi protein trafficking inhibitor) had no effect on the E2-induced
increase in VGSC amplitude, indicating that such trafficking (externalization') of VGSC was not involved. Finally, acute
application of E2 decreased cell adhesion while the specific VGSC blocker tetrodotoxin increased it. Co-application of
E2 and tetrodotoxin inhibited the effect of E2 on cell adhesion, suggesting that the effect of E2 was mainly through VGSC
activity. Pre-treatment of the cells with PKI abolished the effect of E2 on adhesion, consistent with the proposed role of
PKA. Potential implications of the E2-induced non-genomic upregulation of VGSC activity for BCa progression are
discussed.
~20 Citings
254. Methods and compositions using PDE4 inhibitors for treatment and management of hematol. cancers
By Zeldis, Jerome B.; Schafer, Peter H.
Methods of treating, preventing and/or managing hematol. cancers are disclosed. Specific methods encompass the
administration of a PDE4 inhibitor alone or in combination with a second active agent. The invention further relates to
methods of treating leukemias and lymphomas which comprise the administration of a PDE4 inhibitor. Pharmaceutical
compns., single unit dosage forms, and kits suitable for use in methods of the invention are also disclosed. Thus, a
PDE4 inhibitor of the invention is cyclically administered to patients with cancer; cycling therapy involves the
administration of a first agent for a period of time, followed by a rest for a period of time and repeating this sequential
administration; cycling therapy can reduce the development of resistance to one or more of the therapies, avoid or
reduce the side effects of one of the therapies, and/or improves the efficacy of the treatment. In a specific embodiment,
prophylactic or therapeutic agents are administered in a cycle of about four to six weeks, about once or twice every day;
one cycle can comprise the administration of a therapeutic on prophylactic agent for three to four weeks and at least one
week or two weeks of rest; the no. of cycles administered is from about one to about 24 cycles, more typically from about
two to about 16 cycles, and more typically from about four to about eight cycles.
~0 Citings
255. Methods for characterizing molecules in drug discovery and generating a molecule biosensor index
By Fang, Ye; Ferrie, Ann M.; Lahiri, Joydeep; Tran, Elizabeth
Drug discovery is a complex undertaking facing many challenges, not the least of which is a high attrition rate as many
promising candidates prove ineffective or toxic in the clinic owing to a poor understanding of the diseases, and thus the
biol. systems, they target. Therefore, it is broadly agreed that to increase the productivity of drug discovery one needs a
far deeper understanding of the mol. mechanisms of diseases, taking into account the full biol. context of the drug target
and moving beyond individual genes and proteins. The disclosure provides methods, compn., articles, and machines for
characterizing a mol. using a label-free biosensor cellular assay, and performing systems biol. and systems pharmacol.
anal. of mols. including ligands, as well as drug discovery. The disclosure also provides methods, compns., articles, and
machines for producing an index for a given mol., particularly the DMR (dynamic mass redistribution) index, and the
method of using comparative anal. of the indexes of different mols. to det. the mode(s) of action of any mols.
~1 Citing
256. Methods for characterizing molecules in drug discovery and generating a molecule biosensor index
By Fang, Ye; Ferrie, Ann M.; Lahiri, Joydeep; Tran, Elizabeth
Drug discovery is a complex undertaking facing many challenges, not the least of which is a high attrition rate as many
promising candidates prove ineffective or toxic in the clinic owing to a poor understanding of the diseases, and thus the
biol. systems, they target. Therefore, it is broadly agreed that to increase the productivity of drug discovery one needs a
far deeper understanding of the mol. mechanisms of diseases, taking into account the full biol. context of the drug target
and moving beyond individual genes and proteins. The disclosure provides methods, compn., articles, and machines for
characterizing a mol. using a label-free biosensor cellular assay, and performing systems biol. and systems pharmacol.
anal. of mols. including ligands, as well as drug discovery. The disclosure also provides methods, compns., articles, and
machines for producing an index for a given mol., particularly the DMR (dynamic mass redistribution) index, and the
method of using comparative anal. of the indexes of different mols. to det. the mode(s) of action of any mols.
~0 Citings
257. Activated Wnt-beta-catenin signaling in melanoma

By Moon, Randall T.; Chien, Andy J.; Rimm, David L.
The present invention is directed to a method of detg. the prospects for survival of a melanoma patient. This method
involves providing a biol. sample from a patient diagnosed with melanoma, detg. the level of an indicator of Wnt/β-catenin
activation in the sample, comparing the level of the indicator of Wnt/β-catenin activation in the sample against a std. level
of the indicator of Wnt/β-catenin activation correlated to survival of melanoma, and detg. a patient's prospects for survival
of melanoma based on the comparison. The present invention also relates to a method of improving survival of
melanoma patients, decreasing metastases in melanoma patients, decreasing proliferation of cancer cells in melanoma
patients, decreasing melanoma recurrence in melanoma patients, and/or decreasing tumor size in melanoma patients.
This involves selecting melanoma patients and subsequently detg. and monitoring therapy based on their level of Wnt/β-
catenin activation. The selected melanoma patients are treated with a dose of an activator or synergizer of Wnt/β-
catenin based on the patient's level of Wnt/β-catenin, under conditions effective, resp., to improve survival of the selected
melanoma patients, to decrease metastases in the selected melanoma patients, to decrease proliferation of cancer cells
in the selected melanoma patients, to decrease melanoma recurrence in the selected melanoma patients, and/or to
decrease tumor size in the selected melanoma patients.
~1 Citing
258. Differential regulation and roles of urocortins in human adrenal H295R cells
By Kageyama, Kazunori; Hanada, Komaki; Suda, Toshihiro
From Regulatory Peptides (2010), 162(1-3), 18-25. Language: English, Database: CAPLUS,
DOI:10.1016/j.regpep.2010.02.006
Three urocortins (Ucns) are known as members of the corticotropin-releasing factor (CRF) family of peptides and serve
as natural ligands for CRF receptors. Ucn1 and Ucn3 exhibit potent effects on the adrenal system via the CRF receptors.
This study aimed to explore the regulation and roles of Ucns in the adrenal system using human adrenal carcinoma
H295R cells, which express Ucn1, Ucn2, Ucn3, CRF receptor type 1 (CRF1 receptor), and CRF receptor type 2a (CRF2a
receptor) mRNA. Forskolin, which stimulates adenylate cyclase and then increases intracellular cAMP prodn., was
shown to transiently decrease Ucn1 and Ucn2 mRNA levels, but increase Ucns 1-3 mRNA levels in H295R cells.
Steroidogenic acute regulatory protein, Cyp11β1, and Cyp11β2 mRNA levels, and both cortisol and aldosterone
secretions were elevated by Ucn1. Cell viability was reduced by both Ucn1 and Ucn3 via the CRF2 receptor in H295R
cells. Ucn1 and Ucn3 increased the expression of the cAMP-response element binding protein and extracellular signal-
related kinase (ERK) phosphorylations. The ERK and protein kinase A pathways were involved in Ucn3-decreased cell
viability.
~13 Citings
259. Effects of catechin, epicatechin and epigallocatechin gallate on testosterone production in rat leydig cells
By Yu, Po-Ling; Pu, Hsiao-Fung; Chen, Sung-Yun; Wang, Shyi-Wu; Wang, Paulus S.
From Journal of Cellular Biochemistry (2010), 110(2), 333-342. Language: English, Database: CAPLUS,
DOI:10.1002/jcb.22541
Catechins have been reported to have many pharmacol. properties such as the effects of anti-oxidative, anti-
inflammatory, anti-carcinogenic, anti-UV, and redn. of blood pressure as well as glucose and cholesterol levels.
However, the effect of catechins on the reproductive mechanism is still unknown. In the present study, the effects of
catechins on testosterone secretion in rat testicular Leydig cells (LCs) were explored. Both in vivo and in vitro
investigations were performed. Purified LCs were incubated with or without catechin (CCN), epicatechin (EC),
epigallocatechin gallate (EGCG, 10-10-10-8 M) under challenge with human chorionic gonadotropin (hCG, 0.01 IU/mL),
forskolin, SQ22536 (an adenylyl cyclase inhibitor), 8-bromo-adenosine 3':5'-cyclic monophosphate (8-Br-cAMP), A23187
(a calcium ionophore), and nifedipine (10-5 M), resp. To study the effects of catechins on steroidogenesis, steroidogenic
precursors-stimulated testosterone release was examd. The functions of the steroidogenic enzymes including protein
expression of cytochrome P 450 side chain cleavage enzyme (P450scc) and steroidogenic acute regulatory (StAR)
protein were investigated and expressed by Western blotting. Catechins increased plasma testosterone in vivo in male
rats. In vitro, low-dose concn. of catechins increased gonadotropin releasing hormone (GnRH)-stimulated LH release by
anterior pituitary gland and hCG-stimulated testosterone release by LCs of male rats. These results suggested that
catechins stimulated testosterone prodn. by acting on rat LCs via the mechanism of increasing the action of cAMP, but
not P450scc, StAR protein or the activity of intracellular calcium. EC, one of the catechins increased the testosterone
secretion by rat LCs via the enzyme activities of 17β-hydroxysteroid dehydrogenase (17β-HSD).
~8 Citings
260. A role for inflammatory mediators in heterologous desensitization of CysLT1 receptor in human monocytes
By Capra, Valerie; Accomazzo, Maria Rosa; Gardoni, Fabrizio; Barbieri, Silvia; Rovati, G. Enrico
From Journal of Lipid Research (2010), 51(5), 1075-1084. Language: English, Database: CAPLUS,
DOI:10.1194/jlr.M003236
Cysteinyl-leukotrienes (cysteinyl-LT) are rapidly generated at sites of inflammation and, in addn. to their role in asthma,
rhinitis, and other immune disorders, are increasingly regarded as significant inflammatory factors in cancer,
gastrointestinal, cardiovascular diseases. We recently demonstrated that in monocyte/macrophage-like U937 cells,
extracellular nucleotides heterologously desensitize CysLT1 receptor (CysLT1R)-induced Ca2+ transients. Given that
monocytes express a no. of other inflammatory and chemoattractant receptors, this study was aimed at characterizing
transregulation between these different stimuli. We demonstrate that in U937 cells and in primary human monocytes, a
series of inflammatory mediators activating Gi-coupled receptor (FPR1, BLT1) desensitize CysLT1R-induced Ca2+
response unidirectionally through activation of PKC. Conversely, PAF-R, exclusively coupled to Gq, cross-desensitizes
CysLT1R without the apparent involvement of any kinase. Interestingly, Gs-coupled receptors (β2AR, H1/2R, EP2/4R) are
also able to desensitize CysLT1R response through activation of PKA. Heterologous desensitization seems to affect
mostly the Gi-mediated signaling of the CysLT1R. The hierarchy of desensitization among agonists may be important for
leukocyte signal processing at the site of inflammation. Considering that monocytes/macrophages are likely to be the
major source of cysteinyl-LT in many immunol. and inflammatory processes, shedding light on how their receptors are
regulated will certainly help to better understand the role of these cells in orchestrating this complex network of integrated
signals.
~7 Citings
261. Modulation of axon degeneration and therapeutical uses in treating neurological disorders and nervous
system injuries
By Watts, Ryan; Chen, Mark; Lewcock, Joseph Wesley; Pozniak, Christine; Sengupta-Ghosh, Arundhati
The invention relates generally to treatment of neurol. disorders and nervous system injuries. The invention specifically
provides methods of using modulators of particular target proteins to modulate degeneration of neurons or portions
thereof, such as axons.
~6 Citings
262. Isoproterenol and cAMP Block ERK Phosphorylation and Enhance [Ca2+]i Increases and Oxygen
Consumption by Muscarinic Receptor Stimulation in Rat Parotid and Submandibular Acinar Cells
By Soltoff, Stephen P.; Hedden, Lee
DOI:10.1074/jbc.M110.112094
Salivary glands are innervated by sympathetic and parasympathetic neurons, which release neurotransmitters that
promote fluid secretion and exocytosis when they bind to muscarinic and β-adrenergic receptors, resp. Signaling
pathways downstream of these receptors are mainly distinct, but there is cross-talk that affects receptor-dependent
events. Here we report that the β-adrenergic ligand isoproterenol blocks increases in extracellular signal-related kinase
(ERK) phosphorylation, a protein kinase C-dependent event promoted by the muscarinic receptor ligand carbachol in
freshly dispersed rat parotid acinar cells. The inhibitory action of isoproterenol was reproduced by cAMP stimuli
(forskolin) and mimetics (dibutyryl-cAMP, 8-(4-chlorophenylthio)-cAMP), including one highly selective for protein kinase
A (N6-benzoyl-cAMP). In contrast, Epac (exchange proteins directly activated by cAMP)-selective activators did not
mimic the blockade of ERK by isoproterenol, suggesting that inhibition involved protein kinase A. Isoproterenol also
blocked ERK downstream of phorbol 12-myristate 13-acetate and the P2X7 and epidermal growth factor receptors.
Isoproterenol and forskolin blocked MEK phosphorylation, reduced RAF phosphorylation on a stimulatory site (Ser-338),
and increased RAF phosphorylation on an inhibitory site (Ser-259). Inhibitory effects on ERK were also obsd. in freshly
dispersed rat submandibular acinar cells but not in three immortalized/cancer salivary cell lines (Par-C10, HSY, HSG),
indicating significant differences between native cells and cell lines. Notably, in native parotid cells isoproterenol
enhanced the carbachol-promoted increases in [Ca2+]i and oxygen consumption, events that initiate and accompany,
resp., the stimulation of fluid secretion by muscarinic ligands. Thus, isoproterenol produces opposite effects on
prominent events downstream of the muscarinic receptor second messengers diacylglycerol (decrease in ERK
phosphorylation) and inositol trisphosphate (increase in [Ca2+]i and fluid secretion).
~10 Citings
263. Krueppel-like factor 4 is widely expressed in the mouse male and female reproductive tract and responds
as an immediately early gene to activation of the protein kinase A in TM4 sertoli cells
By Godmann, M.; Kosan, C.; Behr, R.
From Reproduction (Bristol, United Kingdom) (2010), 139(4), 771-782. Language: English, Database: CAPLUS,
DOI:10.1530/REP-09-0531
Krueppel-like factor 4 (KLF4) is a zinc finger transcription factor critically involved in cell proliferation, differentiation, and
carcinogenesis. Recently, KLF4 has also been used for the generation of induced pluripotent stem cells. In this study,
we analyzed Klf4 expression in different mouse tissues using northern blot anal. and immunohistochem. Focusing on the
male and female reproductive tract, we showed for the first time that KLF4 is expressed in the epithelia of the murine
uterus and the vagina. In the male reproductive tract, we detected KLF4 in the epithelia of the epididymis, ductus
deferens, coagulating gland, and the penis. As KLF4 is strongly inducible by FSH signaling in Sertoli cells and as this
transcription factor is also involved in Sertoli cell development, we employed the mouse Sertoli cell line TM4 as a model
system to investigate (i) the induction kinetics of Klf4 upon activation of the cAMP/protein kinase A pathway by forskolin
and (ii) the effects of Klf4 induction on TM4 cell cycle progression. Interestingly, Klf4 mRNA and protein were rapidly but
transiently induced, reaching peak levels after 90-120 min and declining to basal levels within 4 h. Compared with the
inducible cAMP early repressor, an immediate early response gene, the induction kinetics of Klf4 is much faster. In
conclusion, Klf4 is an immediate early gene in TM4 cells and its expression in several epithelia of the male and female
reproductive tract suggests an important role of Klf4 in mouse reproductive functions.
~10 Citings
264. Neuropeptide Y Y5 receptor promotes cell growth through extracellular signal-regulated kinase signaling
and Cyclic AMP Inhibition in a Human Breast Cancer Cell Line
By Sheriff, Sulaiman; Ali, Marwan; Yahya, Ayesha; Haider, Khawaja H.; Balasubramaniam, Ambikaipakan; Amlal,
Hassane
From Molecular Cancer Research (2010), 8(4), 604-614. Language: English, Database: CAPLUS, DOI:10.1158/1541-
7786.MCR-09-0301
Overexpression of neuropeptide Y (NPY) and its receptor system has been reported in various types of cancers. NPY
Y5 receptor (Y5R) has been implicated in cell growth and angiogenesis. However, the role of Y5R in breast cancer is
unknown. To identify the role of Y5R in breast cancer, we screened several breast cancer cell lines to examine the
expression of Y5R and its function in breast cancer. All screened cell lines express both Y1 receptor and Y5R except
BT-549, which expresses mainly Y5R. Binding studies showed that NPY, Y5R-selective agonist peptide, and Y5R-
selective antagonist (CGP71683A) displaced 125I-PYY binding in BT-549 cell membranes in a dose-dependent manner.
The displacement studies revealed the presence of two binding sites in Y5R with IC50 values of 29 pmol/L and 531
nmol/L. NPY inhibited forskolin-stimulated cAMP accumulation with an IC50 value of 52 pmol/L. NPY treatment of BT-
549 cells induced extracellular signal-regulated kinase phosphorylation but did not alter intracellular calcium. Y5R
activation stimulates BT-549 cell growth, which is inhibited by CGP71683A, pertussis toxin, and extracellular signal-
regulated kinase blockade. CGP71683A alone induced cell death in a time- and dose-dependent manner in Y5R-
expressing cells. The stimulation of MDA MB-231 cell migration by NPY is inhibited by CGP71683A. Together, our
results suggest that Y5R plays an important role in cancer cell growth and migration and could be a novel therapeutic
target for breast cancer.
~22 Citings
265. Synergizing active compounds for treating inflammation and other conditions
By Bauer, Nyles
Compns. and methods are provided having unexpected anti-inflammatory and other properties based on unprecedented
synergistic effects of administering two or more of (i) an opiate antagonist, partial antagonist, or reverse agonist, (ii) a
cAMP-phosphodiesterase (PDE) inhibitor, (iii) an adenylate cyclase activator, and (iv) a NAD (phosphate) (NAD(P)H)
oxidase inhibitor. Invention embodiments are useful for treating or reducing one or more of tissue inflammation,
inflammation-assocd. cellular proliferation, oxidative stress or neuronal death or dysfunction, or for treating or preventing
gray hair or restoring hair color.
~2 Citings
266. Use of processed rice as an additive in universal stem cell culture medium for use in therapy and protein
production
By Steele, Ann Marie; Smith, Mark Dargan; Boonchuen, Somchai
The invention relates to the use of processed rice such as rice exts. as an additive to a culture medium to maintain and
grow various cells including stem cells and support their manufd. and secreted products including cytokines, chemokines
and growth factors. The medium allows for metabolic enabling or enhancement of measurable parameters within the
cellular population, availability and usage of nutrients to cells, distinct and pos. effect on cellular dynamics (i.e. cellular
proliferation and secretion of manufd. products by the cells), stabilized environment for maintaining conditions for growth
and other cellular and metabolic processes including secretion of manufd. products including signaling factors, enhanced
biol. cellular function of inherent cellular processes, non-interference with normal cellular metabolics (i.e. signaling),
increased protection from toxic effects to the cell and addnl. benefits to the cellular population which in absence of the
additive would be lacking.
~2 Citings
267. Phosphorylation of VACM-1/Cul5 by Protein Kinase A Regulates Its Neddylation and Antiproliferative Effect
By Bradley, Shirley E.; Johnson, Alyssa E.; Le, Isabelle P.; Oosterhouse, Elizabeth; Hledin, Michael P.; Marquez,
Gabriel A.; Burnatowska-Hledin, Maria
DOI:10.1074/jbc.M109.085225
Expression of the VACM-1/cul5 gene in endothelial and in cancer cell lines in vitro inhibits cellular proliferation and
decreases phosphorylation of MAPK. Structure-function anal. of the VACM-1 protein sequence identified consensus
sites specific for phosphorylation by protein kinases A and C (PKA and PKC) and a Nedd8 protein modification site.
Mutations at the PKA-specific site in VACM-1/Cul5 (S730AVACM-1) sequence resulted in increased cellular growth and
the appearance of a Nedd8-modified VACM-1/Cul5. The aim of this study was to examine if PKA-dependent
phosphorylation of VACM-1/Cul5 controls its neddylation status, phosphorylation by PKC, and ultimately growth. Our
results indicate that in vitro transfection of rat adrenal medullary endothelial cells with anti-VACM-1-specific small
interfering RNA oligonucleotides decreases endogenous VACM-1 protein concn. and increases cell growth. Western blot
anal. of cell lysates immunopptd. with an antibody directed against a PKA-specific phosphorylation site and probed with
anti-VACM-1-specific antibody showed that PKA-dependent phosphorylation of VACM-1 protein was decreased in cells
transfected with S730AVACM-1 cDNA when compared with the cytomegalovirus-transfected cells. This change was
assocd. with increased modification of VACM-1 protein by Nedd8. Induction of PKA activity with forskolin reduced
modification of VACM-1 protein by Nedd8. Finally, rat adrenal medullary endothelial cells transfected with S730AVACM-
1/cul5 cDNA and treated with phorbol 12-myristate 13-acetate (10 and 100 nM) to induce PKC activity grew significantly
faster than the control cells. These results suggest that the antiproliferative effect of VACM-1/Cul5 is dependent on its
posttranslational modifications and will help in the design of new anticancer therapeutics that target the Nedd8 pathway.
~7 Citings

268. Prostaglandin E2-induced colonic secretion in patients with and without colorectal neoplasia
By Kaltoft, Nicolai; Tilotta, Maria C.; Witte, Anne-Barbara; Osbak, Philip S.; Poulsen, Steen S.; Bindslev, Niels;
Hansen, Mark B.
From BMC Gastroenterology (2010), 10, No pp. given. Language: English, Database: CAPLUS, DOI:10.1186/1471-
230X-10-9
The pathogenesis for colorectal cancer remains unresolved. A growing body of evidence suggests a direct correlation
between cyclooxygenase enzyme expression, prostaglandin E2 metab. and neoplastic development. Thus further
understanding of the regulation of epithelial functions by prostaglandin E2 is needed. We hypothesized that patients with
colonic neoplasia have altered colonic epithelial ion transport and express functionally different prostanoid receptor levels
in this respect. Patients referred for colonoscopy were included and grouped into patients with and without colorectal
neoplasia. Patients without endoscopic findings of neoplasia served as controls. Biopsy specimens were obtained from
normally appearing mucosa in the sigmoid part of colon. Biopsies were mounted in miniaturized modified Ussing air-
suction chambers. Indomethacin (10 µM), various stimulators and inhibitors of prostanoid receptors and ion transport
were subsequently added to the chamber solns. Electrogenic ion transport parameters (short circuit current and slope
conductance) were recorded. Tissue pathol. and tissue damage before and after expts. was assessed by histol.
Baseline short circuit current and slope conductance did not differ between the two groups. Patients with neoplasia were
significantly more sensitive to indomethacin with a decrease in short circuit current of 15.1 ± 2.6 µA/cm-2 compared to
controls, who showed a decrease of 10.5 ± 2.1 µA/cm-2 (p = 0.027). Stimulation or inhibition with theophylline, ouabain,
bumetanide, forskolin or the EP receptor agonists prostaglandin E2, butaprost, sulprostone and prostaglandin E1 (OH)
did not differ significantly between the two groups. Histol. was with normal findings in both groups. Epithelial
electrogenic transport is more sensitive to indomethacin in normal colonic mucosa from patients with previous or present
colorectal neoplasia compared to colonic mucosa from control patients. Stimulated epithelial electrogenic transport
through individual prostanoid subtype receptors EP1, EP2, EP3, and EP4 is not significantly different between neoplasia
diseased patients and controls. This indicates that increased indomethacin-sensitive mechanisms in colonic mucosa
from neoplasia diseased patients are not related to differences in functional expression of EP receptor subtypes.
~0 Citings
269. Prostaglandin E2 activates cAMP response element-binding protein in glioma cells via a signaling pathway
involving PKA-dependent inhibition of ERK
By Bidwell, Philip; Joh, Kiwon; Leaver, H. Anne; Rizzo, Maria Teresa
From Prostaglandins & Other Lipid Mediators (2010), 91(1-2), 18-29. Language: English, Database: CAPLUS,
DOI:10.1016/j.prostaglandins.2009.12.002
Prostaglandin E2 (PGE2) plays a crit. role in influencing the biol. behavior of tumor cells. We previously demonstrated
that PGE2 stimulates human glioma cell growth via activation of protein kinase A (PKA) type II. This study was
undertaken to further elucidate the intracellular pathways activated by PGE2 downstream to PKA. Stimulation of U87-
MG glioma cells with PGE2 increased phosphorylation of the cyclic-AMP response element (CRE) binding protein CREB
at Ser-133 and CREB-driven transcription in a dose- and time-dependent manner. Expression of dominant CREB
constructs that interfere with CREB phosphorylation at Ser-133 or with its binding to the CRE site markedly decreased
PGE2-induced CREB activation. Inhibition of PKA by H-89 or expression of a dominant neg. PKA construct attenuated
PGE2-induced CREB activation. Moreover, inhibition of PKA type II decreased PGE2-induced CREB-dependent
transcription by 45% compared to vehicle-treated cells. To investigate the involvement of addnl. signaling pathways,
U87-MG cells were pretreated with wortmannin or LY294002 to inhibit the PI3-kinase/AKT pathway. Both inhibitors had
no effect on PGE2-induced CREB phosphorylation and transcriptional activity, suggesting that PGE2 activates CREB in a
PI3-kinase/AKT independent manner. Challenge of U87-MG cells with PGE2, at concns. that induced maximal CREB
activation, or with forskolin inhibited extracellular signal-regulated kinase (ERK) phosphorylation. Pretreatment of U87-
MG cells with the ERK inhibitor PD98059, accentuated ERK inhibition and increased CREB phosphorylation at Ser-133
and CREB-driven transcription stimulated by PGE2, suggesting that inhibition of ERK contributes to PGE2-induced CREB
activation. Inhibition of ERK by PGE2 or by forskolin was rescued by treatment of cells with H-89 or by the dominant neg.
PKA construct. Moreover, PGE2 or forskolin inhibited phosphorylation of Raf-1 phosphorylation at Ser-338. Challenge of
U87-MG cells with 11-deoxy-PGE1 increased CREB-driven transcription and stimulated cell growth, while other PGE2
analogs had no effect. Together our results reveal a novel signaling pathway whereby PGE2 signals through PKA to
inhibit ERK and increase CREB transcriptional activity.
~16 Citings
270. Protein kinase A and regulation of neonatal Nav1.5 expression in human breast cancer cells: Activity-
dependent positive feedback and cellular migration
By Chioni, Athina-Myrto; Shao, Dongmin; Grose, Richard; Djamgoz, Mustafa B. A.
Voltage-gated Na+channels (VGSCs) are expressed in excitable cells (e.g. neurons and muscles), as well as in some
classically non-excitable' cells (e.g. fibroblasts), and in carcinomas. In general, functional expression of VGSCs in
plasma membrane (PM) is hierarchical and dynamic. Previously, we have shown that an activity-dependent pos.
feedback mechanism involving cAMP-dependent protein kinase A (PKA) plays a significant role in upregulation of
VGSCs in strongly metastatic rat prostate cancer Mat-LyLu cells expressing Nav1.7. Here, we investigated the possible
role of PKA in VGSC regulation and its functional consequences in strongly metastatic human breast cancer (BCa) MDA-
MB-231 cells, where the neonatal splice form of Nav1.5 (nNav1.5) is the predominant VGSC present. Treatment with the
PKA activator forskolin for 24 h increased mRNA and PM protein levels of nNav1.5, without changing the total VGSC
protein level. Opposite effects were obtained by application of the PKA inhibitor KT5720 or the highly specific VGSC
blocker tetrodotoxin (TTX), the latter implying activity-dependent upregulation. We tested the possibility, therefore, that
the activity dependence of VGSC (nNav1.5) expression involved PKA. Indeed, TTX pretreatment reduced the level of
phosphorylated PKA and eliminated basal and PKA-stimulated cellular migration. These data suggested that activity-
dependent pos. feedback mediated by PKA plays an important role in the functional expression of nNav1.5 in BCa, and
in turn, this enhances the cells' metastatic potential.
~15 Citings
271. Stimulation of p53 transactivation ability by nicastrin in mouse fibroblasts

By Koshida, Toshifumi; Kitagawa, Motoo; Iwase, Katsuro; Takiguchi, Masaki; Hiwasa, Takaki
From Scholarly Research Exchange (2010), No pp. given. Language: English, Database: CAPLUS,
DOI:10.3814/2010/606391
Nicastrin (NCSTN), a component of the γ-secretase complex, is involved in the p53-dependent apoptosis of neurons,
although its mechanism remains unclear. We analyzed the effects of NCSTN transfection on the transactivity of p53 in
ras-NIH3T3 mouse fibroblasts with a luciferase assay. Luciferase activity was elevated after transfection, suggesting the
stimulation of p53 transactivation ability. In addn., the protein levels of endogenous mouse p53 and transfected human
p53 increased. The effects of NCSTN appeared to be independent of γ-secretase activity because it was not inhibited by
the γ-secretase inhibitor DAPT. The functional domains of NCSTN were further examd. with NCSTN deletion mutants.
Activation of the p53-responsive promoter was completely diminished in a NCSTN mutant lacking the amino acid
residues between 306 and 360. Since this domain is a γ-secretase-substrate-recognition site, the activation of p53 by
NCSTN may be mediated by γ-secretase-substrate-like mols.
~0 Citings
272. Forskolin, a Hedgehog signal inhibitor, inhibits cell proliferation and induces apoptosis in pediatric tumor
cell lines
By Yamanaka, Hiroaki; Oue, Takaharu; Uehara, Shuichiro; Fukuzawa, Masahiro
From Molecular Medicine Reports (2010), 3(1), 133-139. Language: English, Database: CAPLUS,
DOI:10.3892/mmr_00000230
The Hedgehog (Hh) signaling pathway regulates the development of many organs in mammals. Recent studies have
indicated that the activation of the Hh signaling pathway contributes to the growth of various adult cancers. However,
little is known about its role in the development of pediatric malignancies. The present study was undertaken to examine
the expression and functional involvement of Hh signal transcription factors in pediatric tumor cells to det. their potential
as therapeutic targets. We utilized real-time RT-PCR to investigate the expression of Glioma-assocd. oncogene
homolog 1 (Gli1) in various pediatric tumor cell lines, including rhabdomyosarcoma, neuroblastoma and hepatoblastoma.
The mRNA expression of Gli1 was markedly increased in rhabdomyosarcoma (RMS-YM, RD, RH30) cell lines, and
moderately increased in neuroblastoma (NB19) and hepatoblastoma (Huh6) cell lines. The proliferation of these cell
lines was dose dependently inhibited by Forskolin, a specific Hh signal inhibitor. In addn., Forskolin-induced growth
suppression was assocd. with the down-regulation of C-Myc. Moreover, the blockade of Hh signaling with Forskolin
enhanced cell apoptosis in a dose-dependent manner. These results demonstrated that Hh signal activation frequently
occurs in neuroblastoma, hepatoblastoma and rhabdomyosarcoma cell lines. The inhibition of Hh signaling suppressed
proliferation and increased apoptosis in these tumor cells. These findings suggest that the Hh signaling pathway plays
an important role in tumorigenesis and is a potential mol. target of new treatment strategies for these pediatric malignant
tumors.
~4 Citings
273. Synergistic interactions between sorafenib and bortezomib in hepatocellular carcinoma involve PP2A-
dependent Akt inactivation
By Chen, Kuen-Feng; Yu, Hui-Chuan; Liu, Tsung-Hao; Lee, Shoei-Sheng; Chen, Pei-Jer; Cheng, Ann-Lii
From Journal of Hepatology (2010), 52(1), 88-95. Language: English, Database: CAPLUS,
DOI:10.1016/j.jhep.2009.10.011
Background & Aims: Previously we reported that Akt inactivation dets. the sensitivity of hepatocellular carcinoma (HCC)
cells to bortezomib. Here we report that combined treatment with sorafenib and bortezomib shows synergistic effects in
HCC. Methods: HCC cell lines (PLC/PRF/5, Huh-7, and Hep3B) were treated with sorafenib and/or bortezomib and
analyzed in terms of apoptosis signal transduction. In vivo efficacy was detd. in nude mice with PLC/PRF/5 xenografts.
Results: Pretreatment with sorafenib enhanced bortezomib-induced apoptotic cell death by restoring bortezomib's ability
to inactivate Akt in PLC/PRF/5 cells. Knocking down Akt1 by RNA-interference overcame apoptotic resistance to
bortezomib in PLC/PRF/5 cells and ectopic expression of active Akt in HCC cells abolished the bortezomib sensitizing
effect of sorafenib, indicating Akt inactivation plays a key role in mediating the combinational effects. Moreover, okadaic
acid, a protein phosphatase 2A (PP2A) inhibitor, reversed down-regulation of phospho-Akt (P-Akt) expression induced by
co-treatment with sorafenib and bortezomib, and 1, 9 di-deoxy-forskolin, a PP2A agonist, restored bortezomib's effect on
P-Akt and apoptosis. Importantly, silencing of PP2A by RNA-interference reduced the apoptotic effect induced by
sorafenib-bortezomib co-treatment, indicating that PP2A is indispensable for mediating the effects of these drugs.
Notably, sorafenib with bortezomib increased PP2A activity in PLC/PRF/5 cells without altering protein levels of PP2A
complex or the interaction between PP2A and Akt proteins. Finally, sorafenib plus bortezomib significantly suppressed
PLC/PRF/5 xenograft tumor growth, down-regulated P-Akt expression, and up-regulated PP2A activity. Conclusions:
The combination of sorafenib and bortezomib shows synergy in HCC through PP2A-dependent Akt inactivation.
~39 Citings
274. Gastrin Induces Nuclear Export and Proteasome Degradation of Menin in Enteric Glial Cells
By Sundaresan Sinju; Meininger Cameron A; Kang Anthony J; Photenhauer Amanda L; Hayes Michael M; Ding Lin;
Sahoo Nirakar; Xu Haoxing; Grembecka Jolanta; Cierpicki Tomasz; et al
From Gastroenterology (2017), , Language: English, Database: MEDLINE
BACKGROUND & AIMS: The multiple endocrine neoplasia, type 1 (MEN1) locus encodes the nuclear protein and
tumor suppressor menin. MEN1 mutations frequently cause neuroendocrine tumors (NETs) such as gastrinomas,
characterized by their predominant duodenal location and local metastasis at time of diagnosis. Diffuse gastrin cell
hyperplasia precedes the appearance of MEN1 gastrinomas, which develop within submucosal Brunner's glands. We
investigated how menin regulates expression of the gastrin gene and induces generation of submucosal gastrin-
expressing cell hyperplasia. METHODS: Primary enteric glial cultures were generated from the
VillinCre:Men(1FL/FL):Sst(-/-) mice or C57BL/6 mice (controls), with or without inhibition of gastric acid by omeprazole.
Primary enteric glial cells from VillinCre:Men1FL/FL:Sst(+/+) mice were incubated with gastrin and separated into
nuclear and cytoplasmic fractions. Cells were incubated with forskolin and H89 to activate or inhibit protein kinase A (a
family of enzymes whose activity depends on cellular levels of cyclic AMP). Gastrin was measured in blood, tissue,
and cell cultures using an ELISA. Immunoprecipitation with menin or ubiquitin was used to demonstrate post-
translational modification of menin. Primary glial cells were incubated with leptomycin b and MG132 to block nuclear
export and proteasome activity, respectively. We obtained human duodenal, lymph node, and pancreatic gastrinoma
samples, collected from patients who underwent surgery from 1996 through 2007 in the United States or the United
Kingdom. RESULTS: Enteric glial cells that stained positive for glial fibrillary acidic protein (GFAP+) expressed gastrin
de novo through a mechanism that required PKA. Gastrin-induced nuclear export of menin via cholecystokinin B
receptor (CCKBR)-mediated activation of PKA. Once exported from the nucleus, menin was ubiquitinated and
degraded by the proteasome. GFAP and other markers of enteric glial cells, e.g., p75 and S100B, colocalized with
gastrin in human duodenal gastrinomas. CONCLUSIONS: MEN1-associated gastrinomas, which develop in the
submucosa, might arise from enteric glial cells through hormone-dependent PKA signaling. This pathway disrupts
nuclear menin function, leading to hypergastrinemia and associated sequelae.
~0 Citings
Copyright © 2017 U.S. National Library of Medicine.
275. Effects of sorafenib and an adenylyl cyclase activator on in vitro growth of well-differentiated thyroid
cancer cells
By Sawa Aya; Chiba Tomohiro; Ishii Jun; Yamamoto Hiroyuki; Kamma Hiroshi; Sawa Aya; Yamamoto Hiroyuki; Hara
Hisato
From Endocrine journal (2017), , Language: English, Database: MEDLINE
Well-differentiated thyroid carcinomas have driver mutations involving growth factor receptor-tyrosine kinases (RTKs)
or their intracellular signaling pathway, that is, the mitogen-activated protein kinase (MAPK) pathway. Sorafenib is a
multikinase inhibitor of RTKs and the MAPK pathway and has recently been used for the treatment of unresectable
well-differentiated thyroid carcinoma. In normal thyroid follicular cells, stimulation of the thyroid-stimulating hormone
(TSH) receptor activates the cyclic adenosine monophosphate (cAMP) pathway and promotes cell growth as well as
hormonal secretion. However, an adenylyl cyclase (AC) activator, forskolin, has been reported to suppress the growth
of thyroid carcinoma cells. To clarify the roles of the MAPK and cAMP pathways in proliferation of well-differentiated
thyroid carcinoma cells, we compared the effects of sorafenib and forskolin in in vitro models. Sorafenib inhibited
constitutive activation of the MAPK pathway, cyclin-dependent kinase 4 (CDK4), and phosphorylated retinoblastoma
protein (RB) in 3 well-differentiated carcinoma cell lines, but it did not show sufficiently effective suppression of cell
growth. Forskolin significantly suppressed the growth of all 3 cell lines and also activated the cAMP pathway and
inhibited expression of cyclin D1. Our results suggest that activation of the cAMP pathway could be more potent than
activation of the MAPK pathway in suppressing proliferation of well-differentiated thyroid cancer cells. We postulate
that the AC activator suppresses growth of thyroid carcinoma cells through undetermined mechanisms.
~0 Citings
276. Synthesis of novel forskolin isoxazole derivatives with potent anti-cancer activity against breast cancer cell
lines
By Burra Srinivas; Rao Ch Prasad; Vijay Kumar P; Voora Vani; Kancha Rama Krishna; David Krupadanam G L
From Bioorganic & medicinal chemistry letters (2017), 27(18), 4314-4318, Language: English, Database: MEDLINE
Forskolin C1-isoxazole derivatives (3,5-regioisomers) (11a-e, 14, 15a-h and 15, 16a-g) were synthesized
regioselectively by adopting 1,3-dipolar cycloadditions. These derivatives were tested using estrogen receptor positive
breast cancer cell lines MCF-7 and BT-474. Majority of the compounds exhibited activity against the p53-positive
MCF-7 breast cancer cells but not against the p53-negative BT-474 breast cancer cells. Among forskolin derivatives,
compounds 11a, 11c, 14a, 14f, 14g, 14h, 15b, 16g and 17b exhibited higher anti-cancer activity against MCF-7 cell line
with an IC50≤1µM. The derivative 14f exhibited highest activity in both p53-positive (MCF-7) and p53-negative (BT-
474) breast cancer cell lines with an IC50 of 0.5µM.
~0 Citings
277. Antihelminthic drug niclosamide inhibits CIP2A and reactivates tumor suppressor protein phosphatase 2A
in non-small cell lung cancer cells
By Kim Myeong-Ok; Choe Min Ho; Yoon Yi Na; Ahn Jiyeon; Hwang Sang-Gu; Yoo Minjin; Jung Kwan-Young; An
Sungkwan; Oh Jeong Su; Kim Jae-Sung
From Biochemical pharmacology (2017), , Language: English, Database: MEDLINE
Protein phosphatase 2A (PP2A) is a critical tumor suppressor complex responsible for the inactivation of various
oncogenes. Recently, PP2A reactivation has emerged asan anticancer strategy. Cancerous inhibitor of protein
phosphatase 2A (CIP2A), an endogenous inhibitor of PP2A, is upregulated in many cancer cells, including non-small
cell lung cancer (NSCLC) cells. We demonstrated that the antihelminthic drug niclosamide inhibited the expression of
CIP2A and reactivated the tumor suppressor PP2A in NSCLC cells. We performed a drug-repurposing screen and
identified niclosamide asa CIP2A suppressor in NSCLC cells. Niclosamide inhibited cell proliferation, colony formation,
and tumor sphere formation, and induced mitochondrial dysfunction through increased mitochondrial ROS production
in NSCLC cells; however, these effects were rescued by CIP2A overexpression, which indicated that the antitumor
activity of niclosamide was dependent on CIP2A. We found that niclosamide increased PP2A activity through CIP2A
inhibition, which reduced the phosphorylation of several oncogenic proteins. Moreover, we found that a niclosamide
analog inhibited CIP2A expression and increased PP2A activity in several types of NSCLC cells. Finally, we showed
that other well-known PP2A activators, including forskolin and FTY720, did not inhibit CIP2A and that their activities
were not dependent on CIP2A. Collectively, our data suggested that niclosamide effectively suppressed CIP2A
expression and subsequently activated PP2A in NSCLC cells. This provided strong evidence for the potential use of
niclosamide asa PP2A-activating drug in the clinical treatment of NSCLC.
~0 Citings

278. Differential proteome expression analysis of androgen-dependent and -independent pathways in LNCaP
prostate cancer cells
By Cha Seho; Seok Jun Ryeong; Shin Dong Hoon; Myung Jae Kyung
From Experimental cell research (2017), 359(1), 215-225, Language: English, Database: MEDLINE
Prostate cancer (PC) is one of the leading causes of cancer death in men. It commonly develops in older males, but
the number of younger men diagnosed with the disease has increased in recent years. Hormone therapies, such as
chemical and surgical methods that inhibit androgen synthesis or androgen receptor (AR) activation, have been used
for advanced disease. However, castration-resistant PC (CRPC), which exhibits androgen-independent mechanisms
for activating AR, develops after a few years of such treatment and no therapy is available. In this study, we examined
differences in the proteomes associated with the androgen-dependent (DHT treatment) and -independent (FSK,
forskolin treatment) AR signaling conditions in LNCaP prostate cancer cells. Moreover, we used EPI-001, which
inhibits AR-mediated transcriptional activity, to examine whether the observed differences in protein expression were
directly affected by AR-mediated mechanisms. A total of 213 protein spots were matched in our 2-dimensional gel
electrophoresis (2DE) analysis and 8 proteins with significant expression changes in our 5 different treatment groups
were identified by mass spectrometry. Among these proteins, expression levels of PEPCK-M, catalase, tubulin alpha
chain, alpha-enolase, and endoplasmic reticulum resident protein 29 were significantly altered by DHT and the levels
of HSP 90 and EF-Tu were changed by FSK. These changes were blocked by EPI-001 in DHT-regulated proteins,
PEPCK-M, catalase, and tubulin alpha chain and FSK-regulated EF-Tu protein. The results from our
immunohistochemical analysis using in vivo xenograft models were consistent with the 2DE data. We therefore report
the first identification of differences in proteins that are significantly regulated under androgen-dependent and -
independent AR signaling conditions. Our findings could suggest a possible molecular mechanism through which AR
is activated in the absence and/or presence of androgen, and might help explain the inhibitory action of EPI-001 on
CRPC.
~0 Citings
279. Assay of Calcium Transients and Synapses in Rat Hippocampal Neurons by Kinetic Image Cytometry and
High-Content Analysis: An In Vitro Model System for Postchemotherapy Cognitive Impairment
By McDonough Patrick M; Basa Ranor C B; Price Jeffrey H; Prigozhina Natalie L; Price Jeffrey H
From Assay and drug development technologies (2017), 15(5), 220-236, Language: English, Database: MEDLINE
Postchemotherapy cognitive impairment (PCCI) is commonly exhibited by cancer patients treated with a variety of
chemotherapeutic agents, including the endocrine disruptor tamoxifen (TAM). The etiology of PCCI is poorly
understood. Our goal was to develop high-throughput assay methods to test the effects of chemicals on neuronal
function applicable to PCCI. Rat hippocampal neurons (RHNs) were plated in 96- or 384-well dishes and exposed to
test compounds (forskolin [FSK], 17β-estradiol [ES]), TAM or fulvestrant [FUL], aka ICI 182,780) for 6-14 days. Kinetic
Image Cytometry® (KIC®) methods were developed to quantify spontaneously occurring intracellular calcium
transients representing the activity of the neurons, and high-content analysis (HCA) methods were developed to
quantify the expression, colocalization, and puncta formed by synaptic proteins (postsynaptic density protein-95 [PSD-
95] and presynaptic protein Synapsin-1 [Syn-1]). As quantified by KIC, FSK increased the occurrence and
synchronization of the calcium transients indicating stimulatory effects on RHN activity, whereas TAM had inhibitory
effects. As quantified by HCA, FSK also increased PSD-95 puncta and PSD-95:Syn-1 colocalization, whereas ES
increased the puncta of both PSD-95 and Syn-1 with little effect on colocalization. The estrogen receptor antagonist
FUL also increased PSD-95 puncta. In contrast, TAM reduced Syn-1 and PSD-95:Syn-1 colocalization, consistent with
its inhibitory effects on the calcium transients. Thus TAM reduced activity and synapse formation by the RHNs, which
may relate to the ability of this agent to cause PCCI. The results illustrate that KIC and HCA can be used to quantify
neurotoxic and neuroprotective effects of chemicals in RHNs to investigate mechanisms and potential therapeutics for
PCCI.
~0 Citings
280. Smurf1 regulates lung cancer cell growth and migration through interaction with and ubiquitination of
PIPKIγ
By Li H; Ge X; Li H; Li H; Xiao N; Wang Y; Chen Y; Pan W; Liu D; Li S; et al
From Oncogene (2017), , Language: English, Database: MEDLINE
Type Iγ phosphatidylinositol phosphate kinase (PIPKIγ), a phospholipid kinase generating PIP2, is positively expressed
in breast cancer tissues, which correlates intimately with the progression of patients. However, little is known about the
expression level of PIPKIγ in patients with other cancer types as well as their underlying regulation mechanisms. Here,
we report that PIPKIγ is highly expressed in lung cancer tissues and its expression level is critical for lung cancer cell
proliferation, which may serve as a prognostic marker for lung cancer patients. Meanwhile, we show that E3 ubiquitin
ligase Smurf1 directly interacts with PIPKIγ and targets PIPKIγ for ubiquitination and degradation in lung cancer cells.
Also, we discover that Smurf1 directly binds to the kinase domain of PIPKIγ via its C2 domain while Lysine 255 in
PIPKIγ acts as the major ubiquitin acceptor site for Smurf1. In addition, we demonstrate that the phosphorylation
mimicking mutant of Smurf1, Smurf1 T306D, prevents PIPKIγi2 from ubiquitination and subsequent degradation similar
to the effect of forskolin-potentiated cAMP formation, suggesting that Thr306 in Smurf1 is critical for its phosphorylation
by PKA. Moreover, PKA-Smurf1-PIPKIγ signal transduction takes a significant part in lung cancer cell growth and in
vivo tumorigenesis. Thus, we propose that the PKA-Smurf1-PIPKIγ pathway has an important role in pulmonary
tumorigenesis and imposes substantial clinical impact on development of novel diagnostic markers and therapeutic
targets for lung cancer treatment.Oncogene advance online publication, 5 June 2017; doi:10.1038/onc.2017.166.
~0 Citings
281. Catecholamines facilitate VEGF-dependent angiogenesis via β2-adrenoceptor-induced Epac1 and PKA
activation
By Garg Jaspal; Feng Yu-Xi; Jansen Sepp R; Wieland Thomas; Friedrich Julian; Lezoualc'h Frank; Schmidt Martina
From Oncotarget (2017), 8(27), 44732-44748, Language: English, Database: MEDLINE
Chronic stress has been associated with the progression of cancer and antagonists for β-adrenoceptors (βAR) are
regarded as therapeutic option. As they are also used to treat hemangiomas as well as retinopathy of prematurity, a
role of endothelial β2AR in angiogenesis can be envisioned. We therefore investigated the role of β2AR-induced
cAMP formation by analyzing the role of the cAMP effector molecules exchange factor directly activated by cAMP 1
(Epac1) and protein kinase A (PKA) in endothelial cells (EC). Epac1-deficient mice showed a reduced amount of pre-
retinal neovascularizations in the model of oxygen-induced retinopathy, which is predominantly driven by vascular
endothelial growth factor (VEGF). siRNA-mediated knockdown of Epac1 in human umbilical vein EC (HUVEC)
decreased angiogenic sprouting by lowering the expression of the endothelial VEGF-receptor-2 (VEGFR-2).
Conversely, Epac1 activation by β2AR stimulation or the Epac-selective activator cAMP analog 8-p-CPT-2'-O-Me-
cAMP (8-pCPT) increased VEGFR-2 levels and VEGF-dependent sprouting. Similar to Epac1 knockdown, depletion of
the monomeric GTPase Rac1 decreased VEGFR-2 expression. As Epac1 stimulation induces Rac1 activation, Epac1
might regulate VEGFR-2 expression through Rac1. In addition, we found that PKA was also involved in the regulation
of angiogenesis in EC since the adenylyl cyclase (AC) activator forskolin (Fsk), but not 8-pCPT, increased sprouting in
Epac1-depleted HUVEC and this increase was sensitive to a selective synthetic peptide PKA inhibitor. In accordance,
β2AR- and AC-activation, but not Epac1 stimulation increased VEGF secretion in HUVEC.Our data indicate that high
levels of catecholamines, which occur during chronic stress, prime the endothelium for angiogenesis through a β2AR-
mediated increase in endothelial VEGFR-2 expression and VEGF secretion.
~0 Citings
282. β2 adrenergic agonist suppresses eosinophil-induced epithelial-to-mesenchymal transition of bronchial

epithelial cells
By Kainuma Keigo; Kuwabara Yu; Hosoki Koa; Nagao Mizuho; Fujisawa Takao; Kainuma Keigo; D'Alessandro-
Gabazza Corina N; Toda Masaaki; Yasuma Taro; Nishihama Kota; et al
From Respiratory research (2017), 18(1), 79, Language: English, Database: MEDLINE
BACKGROUND: Epithelial-mesenchymal transition is currently recognized as an important mechanism for the
increased number of myofibroblasts in cancer and fibrotic diseases. We have already reported that epithelial-
mesenchymal transition is involved in airway remodeling induced by eosinophils. Procaterol is a selective and full β2
adrenergic agonist that is used as a rescue of asthmatic attack inhaler form and orally as a controller. In this study, we
evaluated whether procaterol can suppress epithelial-mesenchymal transition of airway epithelial cells induced by
eosinophils. METHODS: Epithelial-mesenchymal transition was assessed using a co-culture system of human
bronchial epithelial cells and primary human eosinophils or an eosinophilic leukemia cell line. RESULTS: Procaterol
significantly inhibited co-culture associated morphological changes of bronchial epithelial cells, decreased the
expression of vimentin, and increased the expression of E-cadherin compared to control. Butoxamine, a specific β2-
adrenergic antagonist, significantly blocked changes induced by procaterol. In addition, procaterol inhibited the
expression of adhesion molecules induced during the interaction between eosinophils and bronchial epithelial cells,
suggesting the involvement of adhesion molecules in the process of epithelial-mesenchymal transition. Forskolin, a
cyclic adenosine monophosphate-promoting agent, exhibits similar inhibitory activity of procaterol. CONCLUSIONS:
Overall, these observations support the beneficial effect of procaterol on airway remodeling frequently associated with
chronic obstructive pulmonary diseases.
~0 Citings
283. Prostaglandin E2-mediated adenosinergic effects on CD14(+) cells: Self-amplifying immunosuppression in

cancer
By Al-Taei Saly; Salimu Josephine; Spary Lisa K; Clayton Aled; Tabi Zsuzsanna; Lester Jason F
From Oncoimmunology (2017), 6(2), e1268308, Language: English, Database: MEDLINE
CD39 and CD73 are surface-expressed ectonucleotidases that hydrolyze ATP in a highly regulated, serial manner into
ADP, AMP and adenosine. The end product, adenosine, has both tumor-promoting and immunosuppressive effects.
The aim of this study was to determine CD73 expression on immune cells in pleural effusion (PE) in order to have a
better understanding of the immune environment in mesothelioma. PE- or blood-derived CD14(+) cells of
mesothelioma patients and healthy donors were analyzed by flow cytometry for the expression of CD39 and CD73.
CD73-induction was studied by exposure of CD14(+) cells to the soluble fraction of PE (sPE), while the signaling
mechanism, responsible for CD73 induction, by phosphoflow cytometry and receptor-inhibition studies. We observed
CD73 expression on CD14(+) cells in PE but not peripheral blood of mesothelioma patients or healthy donors. CD73
expression was inducible on CD14(+) cells with sPE, cyclic-AMP (cAMP)-inducers (forskolin and prostaglandin-E2
(PGE2)) and adenosine. Inhibition of PGE2 receptors or adenosine A2 receptors blocked CD73-induction by sPE. sPE
treatment triggered protein kinase A and p38 activation. However, signal-transducer and activator of transcription 3
(STAT3)-blocking led to enhanced CD73 expression, demonstrating a hitherto unknown negative control of purinergic
signaling by STAT3 in CD14(+) cells. TNFα production by CD73(+) CD14(+) cells was significantly impaired in the
presence of AMP, confirming immunosuppressive function. Taken together, CD73 expression can be induced by
PGE2, cAMP or adenosine on human CD14(+) cells. We suggest that targeting this autocrine loop is a valid
therapeutic approach in mesothelioma that may also enhance immunotherapy.
~0 Citings
284. Carfilzomib induces leukaemia cell apoptosis via inhibiting ELK1/KIAA1524 (Elk-1/CIP2A) and activating
PP2A not related to proteasome inhibition
By Liu Chun-Yu; Huang Tzu-Ting; Liu Chun-Yu; Tsai Wen-Chun; Huang Tzu-Ting; Ko Po-Shen; Hung Man-Hsin;
Wang Wan-Lun; Liu Chun-Yu; Huang Chun-Teng; et al
From British journal of haematology (2017), 177(5), 726-740, Language: English, Database: MEDLINE
Enhancing the tumour suppressive activity of protein phosphatase 2A (PP2A) has been suggested to be an anti-
leukaemic strategy. KIAA1524 (also termed CIP2A), an oncoprotein inhibiting PP2A, is associated with disease
progression in chronic myeloid leukaemia and may be prognostic in cytogenetically normal acute myeloid leukaemia.
Here we demonstrated that the selective proteasome inhibitor, carfilzomib, induced apoptosis in sensitive primary
leukaemia cells and in sensitive leukaemia cell lines, associated with KIAA1524 protein downregulation, increased
PP2A activity and decreased p-Akt, but not with the proteasome inhibition effect of carfilzomib. Ectopic expression of
KIAA1524, or pretreatment with the PP2A inhibitor, okadaic acid, suppressed carfilzomib-induced apoptosis and
KIAA1524 downregulation in sensitive cells, whereas co-treatment with the PP2A agonist, forskolin, enhanced
carfilzomib-induced apoptosis in resistant cells. Mechanistically, carfilzomib affected KIAA1524 transcription through
disturbing ELK1 (Elk-1) binding to the KIAA1524 promoter. Moreover, the drug sensitivity and mechanism of
carfilzomib in xenograft mouse models correlated well with the effects of carfilzomib on KIAA1524 and p-Akt
expression, as well as PP2A activity. Our data disclosed a novel drug mechanism of carfilzomib in leukaemia cells and
suggests the potential therapeutic implication of KIAA1524 in leukaemia treatment.
~0 Citings
285. Direct growth-inhibitory effects of prostaglandin E2 in pancreatic cancer cells in vitro through an EP4/PKA-
mediated mechanism
By Schmidt Andrea; Sinnett-Smith James; Young Steven; Rozengurt Enrique; Chang Hui-Hua; Hines O Joe; Dawson
David W; Eibl Guido
From Surgery (2017), 161(6), 1570-1578, Language: English, Database: MEDLINE
BACKGROUND: There is strong evidence linking inflammation and the development of pancreatic ductal
adenocarcinoma. Cyclooxygenase-2 (COX-2) and COX-2-derived PGE2 are overexpressed in human and murine
pancreatic ductal adenocarcinoma. Several studies have demonstrated an important role of COX-2-derived PGE2 in
tumor-stroma interactions; however, the direct growth effects of prostaglandin E2 (PGE2) on pancreatic ductal
adenocarcinoma cells is less well defined. Our aim was to investigate the effects of PGE2 on pancreatic ductal
adenocarcinoma cell growth and to characterize the underlying mechanisms. METHODS: Human pancreatic ductal
adenocarcinoma cell lines, Panc-1 and MIA PaCa-2, were treated with PGE2 in varying doses (0-10 µM). Effects on
the phosphorylation of ERK1/2 were evaluated by Western blot. Colony formation was observed for cells treated with
PGE2 for 11 days. DNA synthesis was determined by (3H)-thymidine incorporation assay. Gene expression of E-type
prostaglandin (EP)2/EP4 receptors and their correlation with survival in patients with pancreatic ductal
adenocarcinoma were assessed using the RNA-Seq data set from The Cancer Genome Atlas Research Network.
RESULTS: PGE2 decreased the size and number of colonies in Panc-1 but not MIA PaCa-2 cells. In the Panc-1 cells,
PGE2 activated PKA/CREB and decreased phosphorylation of ERK1/2, which was reversed by an EP4 receptor
antagonist, while an EP2 receptor antagonist had no effect. In contrast, in MIA PaCa-2 cells, PGE2 had no effect on
ERK1/2 phosphorylation. Treatment of both Panc-1 and MIA PaCa-2 cells with forskolin/IBMX decreased ERK1/2
phosphorylation. Finally, PGE2 decreased DNA synthesis only in Panc-1 cells, which was reversed by an EP4
receptor antagonist. In human pancreatic ductal adenocarcinoma, high EP2 and low EP4 gene expression was
correlated to worse median overall survival (15.6 vs 20.8 months, log-rank P = .017). CONCLUSION: Our study
provides evidence that PGE2 can inhibit directly pancreatic ductal adenocarcinoma cell growth through an EP4-
mediated mechanism. Together with our gene expression and survival analysis, this observation suggests a protective
role of EP4 receptors in human pancreatic ductal adenocarcinoma that expresses E-type prostaglandin receptors.
~0 Citings
286. Enhancing circadian clock function in cancer cells inhibits tumor growth
By Kiessling Silke; Beaulieu-Laroche Lou; Blum Ian D; Storch Kai-Florian; Cermakian Nicolas; Kiessling Silke; Storch
Kai-Florian; Cermakian Nicolas; Kiessling Silke; Landgraf Dominic; et al
From BMC biology (2017), 15(1), 13, Language: English, Database: MEDLINE
BACKGROUND: Circadian clocks control cell cycle factors, and circadian disruption promotes cancer. To address
whether enhancing circadian rhythmicity in tumor cells affects cell cycle progression and reduces proliferation, we
compared growth and cell cycle events of B16 melanoma cells and tumors with either a functional or dysfunctional
clock. RESULTS: We found that clock genes were suppressed in B16 cells and tumors, but treatments inducing
circadian rhythmicity, such as dexamethasone, forskolin and heat shock, triggered rhythmic clock and cell cycle gene
expression, which resulted in fewer cells in S phase and more in G1 phase. Accordingly, B16 proliferation in vitro and
tumor growth in vivo was slowed down. Similar effects were observed in human colon carcinoma HCT-116 cells.
Notably, the effects of dexamethasone were not due to an increase in apoptosis nor to an enhancement of immune cell
recruitment to the tumor. Knocking down the essential clock gene Bmal1 in B16 tumors prevented the effects of
dexamethasone on tumor growth and cell cycle events. CONCLUSIONS: Here we demonstrated that the effects of
dexamethasone on cell cycle and tumor growth are mediated by the tumor-intrinsic circadian clock. Thus, our work
reveals that enhancing circadian clock function might represent a novel strategy to control cancer progression.
~1 Citing
287. The GIP/GIPR axis is functionally linked to GH-secretion increase in a significant proportion of gsp(-)
somatotropinomas
By Regazzo D; Albiger N M; Ceccato F; Scaroni C; Losa M; Terreni M R; Vazza G; Occhi G; Emanuelli E; Denaro L
From European journal of endocrinology (2017), 176(5), 543-553, Language: English, Database: MEDLINE
OBJECTIVE: Glucose-dependent insulinotropic polypeptide receptor (GIPR) overexpression has been recently
described in a proportion of gsp(-) somatotropinomas and suggested to be associated with the paradoxical increase of
GH (GH-PI) during an oral glucose load. DESIGN AND METHODS: This study was aimed at linking the GIP/GIPR
pathway to GH secretion in 25 somatotropinomas-derived primary cultures and correlating molecular with clinical
features in acromegalic patients. Given the impairment of the GIP/GIPR axis in acromegaly, an additional aim was to
assess the effect of GH/IGF-1 stimulation on GIP expression in the enteroendocrine cell line STC-1. RESULTS:
Nearly 80% of GIPR-expressing somatotropinomas, all of them negative for gsp mutations, show increased GH
secretion upon GIP stimulation, higher sensitivity to Forskolin but not to somatostatin analogs. Besides increased
frequency of GH-PI, GIPR overexpression does not appear to affect acromegalic patients' clinical features. In STC-1
cells transfected with GIP promoter-driven luciferase vector, IGF-1 but not GH induced dose-dependent increase in
luciferase activity. CONCLUSIONS: We demonstrate that GIPR mediates the GH-PI in a significant proportion of
gsp(-) acromegalic patients. In these cases, the stimulatory effect of IGF-1 on GIP promoter support the hypothesis of
a functional GH/IGF-1/GIP axis. Further studies based on larger cohorts and the development of a stable transgenic
model with inducible GIPR overexpression targeted to pituitary somatotroph lineage will be mandatory to establish the
real role of GIPR in the pathogenesis of somatotropinomas.
~0 Citings
288. Multiple SLC and ABC Transporters Contribute to the Placental Transfer of Entecavir
By Ma Zhiyuan; Yang Xi; Jiang Ting; Bai Mengru; Zheng Caihong; Zeng Su; Sun Dongli; Jiang Huidi
From Drug metabolism and disposition: the biological fate of chemicals (2017), 45(3), 269-278, Language: English,
Database: MEDLINE
Entecavir (ETV), a nucleoside analog with high efficacy against hepatitis B virus, is recommended as a first-line
antiviral drug for the treatment of chronic hepatitis B. However, scant information is available on the use of ETV in
pregnancy. To better understand the safety of ETV in pregnant women, we aimed to demonstrate whether ETV could
permeate placental barrier and the underlying mechanism. Our study showed that small amount of ETV could
permeate across placenta in mice. ETV accumulation in activated or nonactivated BeWo cells (treated with or without
forskolin) was sharply reduced in the presence of 100 µM of adenosine, cytidine, and in Na(+) free medium, indicating
that nucleoside transporters possibly mediate the uptake of ETV. Furthermore, ETV was proved to be a substrate of
concentrative nucleoside transporter (CNT) 2 and CNT3, of organic cation transporter (OCT) 3, and of breast cancer
resistance protein (BCRP) using transfected cells expressing respective transporters. The inhibition of ETV uptake in
primary human trophoblast cells further confirmed that equilibrative nucleoside transporter (ENT) 1/2, CNT2/3, OCT3,
and organic cation/carnitine transporter (OCTN) 2 might be involved in ETV transfer in human placenta. Therefore,
ETV uptake from maternal circulation to trophoblast cells was possibly transported by CNT2/3, ENT1/2, and OCTN2,
whereas ETV efflux from trophoblast cells to fetal circulation was mediated by OCT3, and efflux from trophoblast cells
to maternal circulation might be mediated by BCRP, multidrug resistance-associated protein 2, and P-glycoprotein.
The information obtained in the present study may provide a basis for the use of ETV in pregnancy.
~0 Citings
289. Triple negative breast cancer development can be selectively suppressed by sustaining an elevated level of
cellular cyclic AMP through simultaneously blocking its efflux and decomposition
By Wang Wei; Fang Dongdong; Su Shi-Bing; Huang Shuang; Li Yue; Zhu Jessica Y; Huang Shuang; Ding Han-Fei;
Dong Zheng; Jing Qing; et al
From Oncotarget (2016), 7(52), 87232-87245, Language: English, Database: MEDLINE
Triple negative breast cancer (TNBC) has the highest mortality among all breast cancer types and lack of targeted
therapy is a key factor contributing to its high mortality rate. In this study, we show that 8-bromo-cAMP, a cyclic
adenosine monophosphate (cAMP) analog at high concentration (> 1 mM) selectively suppresses TNBC cell growth.
However, commonly-used cAMP-elevating agents such as adenylyl cyclase activator forskolin and pan
phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine (IBMX) are ineffective. Inability of cAMP elevating agents to
inhibit TNBC cell growth is due to rapid diminution of cellular cAMP through efflux and decomposition. By performing
bioinformatics analyses with publically available gene expression datasets from breast cancer patients/established
breast cancer cell lines and further validating using specific inhibitors/siRNAs, we reveal that multidrug resistance-
associated protein 1/4 (MRP1/4) mediate rapid cAMP efflux while members PDE4 subfamily facilitate cAMP
decomposition. When cAMP clearance is prevented by specific inhibitors, forskolin blocks TNBC's in vitro cell growth
by arresting cell cycle at G1/S phase. Importantly, cocktail of forskolin, MRP inhibitor probenecid and PDE4 inhibitor
rolipram suppresses TNBC in vivo tumor development. This study suggests that a TNBC-targeted therapeutic strategy
can be developed by sustaining an elevated level of cAMP through simultaneously blocking its efflux and
decomposition.
~0 Citings
290. Cyclic AMP (cAMP) confers drug resistance against DNA damaging agents via PKAIA in CML cells
By Xiao Ling-Yi; Kan Wai-Ming
From European journal of pharmacology (2017), 794201-208, Language: English, Database: MEDLINE
Cyclic adenosine monophosphate (cAMP) regulates many vital functions such as metabolism, proliferation,
differentiation and death. Depending on cell types and stimulators, cAMP could either promote or attenuate cell death.
cAMP signal can be transduced by protein kinase A (PKA) and/or exchange protein directly activated by cAMP
(EPAC). In CML cells, cAMP may suppress their proliferation and enhance their differentiation. However, the role of
cAMP on DNA damaging agent toxicity and the mechanism involved has not been studied. In this study, we studied
the effect of cAMP on the sensitivity of CML cells to DNA damaging agents. We observed that forskolin (FSK) and
dibutyryl-cAMP (DBcAMP) decreased cisplatin and etoposide-induced cell death in K562 cells. Moreover, PKA
activator prevented K562 cells from DNA damaging agent-induced cell death while EPAC activator had no effect.
Furthermore, we found that the PKA subtype, PKAIA, was involved in cAMP-attenuated resistance in K562 cells.
Taken together, our results suggest that increased cAMP level confers CML cells to acquire a novel mechanism
against DNA damaging agent toxicity via PKAIA. Thus, PKAIA inhibitor may be helpful in overcoming the resistance to
DNA damaging agents in CML cells.
~0 Citings
291. Adenylyl cyclase 3/adenylyl cyclase-associated protein 1 (CAP1) complex mediates the anti-migratory effect
of forskolin in pancreatic cancer cells
By Quinn Sierra N; Graves Sarai H; Dains-McGahee Clayton; Friedman Emilee M; Hassan Humma; Sabbatini Maria E;
Witkowski Piotr
From Molecular carcinogenesis (2017), 56(4), 1344-1360, Language: English, Database: MEDLINE
Pancreatic cancer is one of the most lethal human malignancies. A better understanding of the intracellular
mechanism of migration and invasion is urgently needed to develop treatment that will suppress metastases and
improve overall survival. Cyclic adenosine monophosphate (cyclic AMP) is a second messenger that has shown to
regulate migration and invasion of pancreatic cancer cells. The rise of cyclic AMP suppressed migration and invasion
of pancreatic ductal adenocarcinoma cells. Cyclic AMP is formed from cytosolic ATP by the enzyme adenylyl cyclase
(AC). There are ten isoforms of ACs; nine are anchored in the plasma membrane and one is soluble. What remains
unknown is the extent to which the expression of transmembrane AC isoforms is both modified in pancreatic cancer
and mediates the inhibitory effect of forskolin on cell motility. Using real-time PCR analysis, ADCY3 was found to be
highly expressed in pancreatic tumor tissues, resulting in a constitutive increase in cyclic AMP levels. On the other
hand, ADCY2 was down-regulated. Migration, invasion, and filopodia formation in two different pancreatic
adenocarcinoma cell lines, HPAC and PANC-1 deficient in AC1 or AC3, were studied. We found that AC3, upon
stimulation with forskolin, enhanced cyclic AMP levels and inhibited cell migration and invasion. Unlikely to be due to a
cytotoxic effect, the inhibitory effects of forskolin involved the quick formation of AC3/adenylyl cyclase-associated
protein 1 (CAP1)/G-actin complex, which inhibited filopodia formation and cell motility. Using Western blotting
analysis, forskolin, through AC3 activation, caused phosphorylation of CREB, but not ERK. The effect of CREB
phosphorylation is likely to be associated with long-term signaling changes. © 2016 Wiley Periodicals, Inc.
~0 Citings
292. The Natural cAMP Elevating Compound Forskolin in Cancer Therapy: Is It Time?
By Sapio Luigi; Illiano Michela; Chiosi Emilio; Spina Annamaria; Naviglio Silvio; Gallo Monica; Naviglio Daniele
From Journal of cellular physiology (2017), 232(5), 922-927, Language: English, Database: MEDLINE
Cancer is a major public health problem and the second leading cause of mortality around the world. Although
continuous advances in the science of oncology and cancer research are now leading to improved outcomes for many
cancer patients, novel cancer treatment options are strongly demanded. Naturally occurring compounds from a variety
of vegetables, fruits, and medicinal plants have been shown to exhibit various anticancer properties in a number of in
vitro and in vivo studies and represent an attractive research area for the development of new therapeutic strategies to
fight cancer. Forskolin is a diterpene produced by the roots of the Indian plant Coleus forskohlii. The natural
compound forskolin has been used for centuries in traditional medicine and its safety has also been documented in
conventional modern medicine. Forskolin directly activates the adenylate cyclase enzyme, that generates cAMP from
ATP, thus, raising intracellular cAMP levels. Notably, cAMP signaling, through the PKA-dependent and/or -
independent pathways, is very relevant to cancer and its targeting has shown a number of antitumor effects, including
the induction of mesenchymal-to-epithelial transition, inhibition of cell growth and migration and enhancement of
sensitivity to conventional antitumor drugs in cancer cells. Here, we describe some features of cAMP signaling that are
relevant to cancer biology and address the state of the art concerning the natural cAMP elevating compound forskolin
and its perspectives as an effective anticancer agent. J. Cell. Physiol. 232: 922-927, 2017. © 2016 Wiley Periodicals,
Inc.
~1 Citing
293. Transcriptional profiling of immortalized and K-ras-transformed mouse fibroblasts upon PKA stimulation by
forskolin in low glucose availability
By Chiaradonna Ferdinando; Ricciardiello Francesca; Palorini Roberta; Pirola Yuri
From Genomics data (2016), 9100-4, Language: English, Database: MEDLINE
Forskolin (FSK) induces activation of protein kinase A (PKA). This activation protects specifically some cancer cells
from death induced by glucose starvation. Cell effects upon FSK treatment prompted us to investigate in detail the
physiological role of PKA in the activation of pro-survival mechanisms in glucose starvation. In this regard we
performed a microarray analysis of normal NIH3T3 and transformed NIH3T3-K-ras mouse fibroblasts cultured at 1 mM
glucose and daily treated or not with 10 µM FSK until 72 h of growth, when the samples were collected. The
microarray is deposited into Gene Expression Omnibus under Series GSE68266. The microarray data revealed that
the activation of PKA regulates the expression of genes involved in metabolic, stress-response and pro-survival
processes, like glutamine metabolism, autophagy and unfolded protein response, preventing cancer cell death in
glucose starvation. Altogether these findings suggest that PKA activation, by inducing a complex transcriptional
program, leads to cancer survival in nutrient stress, a typical feature of developing tumor. These transcriptional data,
identifying this important role of PKA, will be useful to identify novel target in cancer therapy.
~0 Citings
294. Sensitivity of imatinib-resistant T315I BCR-ABL CML to a synergistic combination of ponatinib and forskolin
treatment
By Oaxaca Derrick M; Yang-Reid Sun Ah; Ross Jeremy A; Rodriguez Georgialina; Kirken Robert A; Staniswalis Joan
G
From Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine (2016),
37(9), 12643-12654, Language: English, Database: MEDLINE
Tyrosine kinase inhibitors (TKIs) have dramatically improved the life expectancy of patients suffering from chronic
myeloid leukemia (CML); however, patients will eventually develop resistance to TKI therapy or adverse side effects
due to secondary off-target mechanisms associated with TKIs. CML patients exhibiting TKI resistance are at greater
risk of developing an aggressive and drug-insensitive disease. Drug-resistant CML typically arises in response to
spontaneous mutations within the drug binding sites of the targeted oncoproteins. To better understand the
mechanism of drug resistance in TKI-resistant CML patients, the BCR-ABL transformed cell line KCL22 was grown
with increasing concentrations of imatinib for a period of 6 weeks. Subsequently, a drug-resistant derivative of the
parental KCL22 cell line harboring the T315I gatekeeper mutation was isolated and investigated for TKI drug sensitivity
via multi-agent drug screens. A synergistic combination of ponatinib- and forskolin-reduced cell viability was identified
in this clinically relevant imatinib-resistant CML cell line, which also proved efficacious in other CML cell lines. In
summary, this study provides new insight into the biological underpinnings of BCR-ABL-driven CML and potential
rationale for investigating novel treatment strategies for patients with T315I CML.
~0 Citings
295. 144 GPR133 Promotes Glioblastoma Growth in Hypoxia

By Bayin Nermin Sumru; Frenster Joshua; Kane J Robert; Modrek Aram; Shohdy Nadim; MacNeil Douglas J; Zagzag
David; Placantonakis Dimitris G
From Neurosurgery (2016), 63 Suppl 1158-9, Language: English, Database: MEDLINE
INTRODUCTION: Microenvironmental diversity in glioblastoma (GBM) is exemplified by normoxic hypervascular areas
and hypoxic necrotic regions. GBM stem cells (GSCs) play a central role in tumor growth and therapy resistance.
How GSCs adapt to diverse GBM microenvironments remains an important and unanswered question. METHODS:
Using primary human GBM cultures, we discovered that CD133+ GSCs are metabolically adept at expanding in
hypoxic conditions. Transcriptional analysis indicated that CD133+ GSCs have 17.8 ± 8.8-fold enriched expression of
GPR133 (n = 3 biospecimens), a member of the adhesion family of G-protein-coupled receptors. We used genetic,
biochemical, and computational assays to interrogate the role of GPR133 in GBM. RESULTS: Immunostaining of 12
human GBM biospecimens revealed that GPR133 expression is restricted to hypoxic regions of GBM and not present
in normal brain. To test whether GPR133 expression is regulated by oxygen tension, we subjected GBM cultures to
1% O2 and found that GPR133 transcript was consistently upregulated (n = 5 cultures) in HIF1a-dependent manner.
To elucidate GPR133's role in tumor growth, we used small hairpin RNA-mediated knockdown. GPR133 knockdown
depleted CD133+ GSCs and inhibited tumor sphere formation under both normoxic and hypoxic conditions (P < .05).
GPR133 knockdown also prevented in vivo tumor formation and increased survival of implanted mice (n = 4/group).
CD133+ GSCs have 26.2% ± 12.53% higher cAMP levels than CD133- GBM cells (n = 3). GPR133 knockdown
downregulated cAMP levels to 47.25% ± 27.27% of control (n = 3). Forskolin, which activates adenylate cyclase and
boosts cAMP production, rescued the knockdown phenotype. These findings suggest that GPR133 canonical
signaling is mediated by cAMP. Kaplan-Meier survival analysis of 160 The Cancer Genome Atlas (TCGA) patients
indicated that increased GPR133 mRNA in GBM tumors correlated with poor survival (P = .002). CONCLUSION: Our
results suggest that GPR133 acts promotes GBM growth in hypoxia. We propose that GPR133 represents an
attractive novel therapeutic target in GBM.
~0 Citings
296. Synthetic High-Density Lipoprotein (sHDL) Inhibits Steroid Production in HAC15 Adrenal Cells
By Taylor Matthew J; Sanjanwala Aalok R; Morin Emily E; Rowland-Fisher Elizabeth; Anderson Kyle; Schwendeman
Anna; Rainey William E
From Endocrinology (2016), 157(8), 3122-9, Language: English, Database: MEDLINE
High density lipoprotein (HDL) transported cholesterol represents one of the sources of substrate for adrenal steroid
production. Synthetic HDL (sHDL) particles represent a new therapeutic option to reduce atherosclerotic plaque
burden by increasing cholesterol efflux from macrophage cells. The effects of the sHDL particles on steroidogenic
cells have not been explored. sHDL, specifically ETC-642, was studied in HAC15 adrenocortical cells. Cells were
treated with sHDL, forskolin, 22R-hydroxycholesterol, or pregnenolone. Experiments included time and concentration
response curves, followed by steroid assay. Quantitative real-time RT-PCR was used to study mRNA of 3-hydroxy-3-
methyl-glutaryl-coenzyme A reductase, lanosterol 14-α-methylase, cholesterol side-chain cleavage enzyme, and
steroid acute regulatory protein. Cholesterol assay was performed using cell culture media and cell lipid extracts from
a dose response experiment. sHDL significantly inhibited production of cortisol. Inhibition occurred in a concentration-
and time-dependent manner and in a concentration range of 3µM-50µM. Forskolin (10µM) stimulated cortisol
production was also inhibited. Incubation with 22R-hydroxycholesterol (10µM) and pregnenolone (10µM) increased
cortisol production, which was unaffected by sHDL treatment. sHDL increased transcript levels for the rate-limiting
cholesterol biosynthetic enzyme, 3-hydroxy-3-methyl-glutaryl-coenzyme A reductase. Extracellular cholesterol
assayed in culture media showed a positive correlation with increasing concentration of sHDL, whereas intracellular
cholesterol decreased after treatment with sHDL. The current study suggests that sHDL inhibits HAC15 adrenal cell
steroid production by efflux of cholesterol, leading to an overall decrease in steroid production and an adaptive rise in
adrenal cholesterol biosynthesis.
~0 Citings
297. A CIEF-LIF method for simultaneous analysis of multiple protein kinases and screening of inhibitors
By Xiao Lixia; Lin Lihua; Liu Shengquan; Yao Shouzhuo
From Electrophoresis (2016), 37(14), 2075-82, Language: English, Database: MEDLINE
Here, a CIEF-LIF method for multiple protein kinase simultaneous analysis and inhibitors throughput screening with
fast rate and low cost is presented. Comparing with CZE, CIEF-LIF exhibited great focusing ability and high separation
efficiency for substrate and phosphorylated peptides, and is applicable for multiple kinases simultaneous analysis
regardless of their substrate peptides compositions and charge statuses. Thus, highly sensitive analysis for cyclic
adenosine 3', 5'-monophosphate-dependent protein kinase (PKA) and cyclin-dependent kinase 1 (CDK1) was
achieved in CIEF-LIF analysis with detection sensitivity up to 1.25 mU/µL and 0.4 mU/µL, respectively, two magnitudes
higher than that of CZE and comparable with that in nanomaterials or green fluorescent protein-based kinase assay.
Moreover, the inhibition effect of inhibitors on multiple kinases could be simultaneously readout in a single
electrophoretic run, with half maximal inhibitory concentration of H-89 for PKA and Ro-3306 for CDK1 calculated as
37.0 and 35.9 nM, respectively, consistent with literatures reported. The CIEF-LIF also exhibited strong anti-
interference ability in human breast cancer cell lysates analysis and simulators such as forskolin and 3-isobutyl-1-
methylxantine assessment. Therefore, CIEF-LIF is desirable for future biological application and clinical diagnostics
and drug discovery.
~0 Citings
298. Extending prematuration with cAMP modulators enhances the cumulus contribution to oocyte antioxidant
defence and oocyte quality via gap junctions
By Li H J; Sutton-McDowall M L; Thompson J G; Wang X; Sugimura S; Gilchrist R B
From Human reproduction (Oxford, England) (2016), 31(4), 810-21, Language: English, Database: MEDLINE
STUDY QUESTION: Can bovine oocyte antioxidant defence and oocyte quality be improved by extending the duration
of pre-in vitro maturation (IVM) with cyclic adenosine mono-phosphate (cAMP) modulators? SUMMARY ANSWER:
Lengthening the duration of cAMP-modulated pre-IVM elevates intra-oocyte reduced glutathione (GSH) content and
reduces hydrogen peroxide (H2O2) via increased cumulus cell-oocyte gap-junctional communication (GJC),
associated with an improvement in subsequent embryo development and quality. WHAT IS KNOWN ALREADY:
Oocytes are susceptible to oxidative stress and the oocyte's most important antioxidant glutathione is supplied, at least
in part, by cumulus cells. A temporary inhibition of spontaneous meiotic resumption in oocytes can be achieved by
preventing a fall in cAMP, and cyclic AMP-modulated pre-IVM maintains cumulus-oocyte GJC and improves
subsequent embryo development. STUDY DESIGN, SIZE, DURATION: This study consisted of a series of 10
experiments using bovine oocytes in vitro, each with multiple replicates. A range of pre-IVM durations were examined
as the key study treatments which were compared with a control. The study was designed to examine if one of the
oocyte's major antioxidant defences can be enhanced by pre-IVM with cAMP modulators, and to examine the
contribution of cumulus-oocyte GJC on these processes. PARTICIPANTS/MATERIALS, SETTING, METHODS:
Immature bovine cumulus-oocyte complexes were treated in vitro without (control) or with the cAMP modulators; 100
µM forskolin (FSK) and 500 µM 3-isobutyl-1-methyxanthine (IBMX), for 0, 2, 4 or 6 h (pre-IVM phase) prior to IVM.
Oocyte developmental competence was assessed by embryo development and quality post-IVM/IVF. Cumulus-oocyte
GJC, intra-oocyte GSH and H2O2 were quantified at various time points during pre-IVM and IVM, in the presence and
the absence of functional inhibitors: carbenoxolone (CBX) to block GJC and buthionine sulfoximide (BSO) to inhibit
glutathione synthesis. MAIN RESULTS AND THE ROLE OF CHANCE: Pre-IVM with FSK + IBMX increased
subsequent blastocyst formation rate and quality compared with standard IVM (P < 0.05), regardless of pre-IVM
duration. The final blastocyst yields (proportion of blastocysts/immature oocyte) were 26.3% for the control, compared
with 39.2, 35.2 and 34.2%, for the 2, 4 and 6 h pre-IVM FSK + IBMX treatments, respectively. In contrast to standard
IVM (control), pre-IVM with cAMP modulators maintained open gap junctions between cumulus cells and oocytes for
the duration (6 h) of pre-IVM examined, and persisted for a further 8 h in the IVM phase. Cyclic AMP-modulated pre-
IVM increased intra-oocyte GSH levels at the completion of both pre-IVM and IVM, in a pre-IVM duration-dependent
manner (P < 0.05), which was ablated when GJC was blocked using CBX (P < 0.05). By 4 h of pre-IVM treatment with
cAMP modulators, oocyte H2O2 levels were reduced compared the control (P < 0.05), although this beneficial effect
was lost when oocytes were co-treated with BSO. Inhibiting glutathione synthesis with BSO during pre-IVM ablated
any positive benefits of cAMP-mediated pre-IVM on oocyte developmental competence (P < 0.01). LIMITATIONS,
REASONS FOR CAUTION: It is unclear if the improvement in oocyte antioxidant defence and developmental
competence reported here is due to direct transfer of total and/or reduced glutathione from cumulus cells to the oocyte
via gap junctions, or whether a GSH synthesis signal and/or amino acid substrates are supplied to the oocyte via gap
junctions. Embryo transfer experiments are required to determine if the cAMP-mediated improvement in blastocyst
rates leads to improved live birth rates. WIDER IMPLICATIONS OF THE FINDINGS: IVM offers significant benefits to
infertile and cancer patients and has the potential to significantly alter ART practice, if IVM efficiency in embryo
production could be improved closer to that of conventional IVF (using ovarian hyperstimulation). Pre-IVM with cAMP
modulators is a simple and reliable means to improve IVM outcomes. STUDY FUNDING/COMPETING INTERESTS:
This work was supported by grants and fellowships from the National Health and Medical Research Council of
Australia (1007551, 627007, 1008137, 1023210) and by scholarships from the Chinese Scholarship Council (CSC)
awarded to H.J.L. and the Japanese Society for the Promotion of Science Postdoctoral Fellowship for Research
Abroad awarded to S.S. The Fluoview FV10i confocal microscope was purchased as part of the Sensing Technologies
for Advanced Reproductive Research (STARR) facility, funded by the South Australian Premier's Science and
Research Fund. We acknowledge partial support from the Australian Research Council Centre of Excellence for
Nanoscale BioPhotonics (CE140100003). We declare that there is no conflict of interest that could be perceived as
prejudicing the impartiality of the research reported.
~3 Citings
299. Dynamic changes in protein interaction between AKAP95 and Cx43 during cell cycle progression of A549
cells
By Chen Xiaoxuan; Zhuang Wenxin; Teng Bogang; Wang Su; Liang Fengchao; Ma Dan; Zhang Suhui; Zou Xuan;
Yang Wei; Zhang Yongxing; et al
From Scientific reports (2016), 621224, Language: English, Database: MEDLINE
Here we show that A-kinase anchoring protein 95 (AKAP95) and connexin 43 (Cx43) dynamically interact during cell
cycle progression of lung cancer A549 cells. Interaction between AKAP95 and Cx43 at different cell cycle phases was
examined by tandem mass spectrometry(MS/MS), confocal immunofluorescence microscopy, Western blot, and co-
immunoprecipitation(Co-IP). Over the course of a complete cell cycle, interaction between AKAP95 and Cx43
occurred in two stages: binding stage from late G1 to metaphase, and separating stage from anaphase to late G1. The
binding stage was further subdivided into complex binding to DNA in interphase and complex separating from DNA in
metaphase. In late G1, Cx43 translocated to the nucleus via AKAP95; in anaphase, Cx43 separated from AKAP95
and aggregated between two daughter nuclei. In telophase, Cx43 aggregated at the membrane of the cleavage
furrow. After mitosis, Cx43 was absent from the furrow membrane and was located in the cytoplasm. Binding between
AKAP95 and Cx43 was reduced by N-(2-[P-Bromocinnamylamino]-ethyl)-5-isoquinolinesulfonmide (H89) treatment
and enhanced by Forskolin. dynamic interaction between AKAP95 and Cx43 varies with cell cycle progression to
regulate multiple biological processes.
~0 Citings
300. Novel effects of FTY720 on perinuclear reorganization of keratin network induced by

sphingosylphosphorylcholine: Involvement of protein phosphatase 2A and G-protein-coupled receptor-12
By Park Mi Kyung; Kim Hyun Ji; Kim Eun Ji; Kim So Yeon; Kang Gyeoung Jin; Byun Hyun Jung; Park Soyeun; Kim
Sang Hee; Lee Ho; Lee Chang Hoon
From European journal of pharmacology (2016), 77586-95, Language: English, Database: MEDLINE
Sphingosylphosphorylcholine (SPC) evokes perinuclear reorganization of keratin 8 (K8) filaments and regulates the
viscoelasticity of metastatic cancer cells leading to enhanced migration. Few studies have addressed the compounds
modulating the viscoelasticity of metastatic cancer cells. We studied the effects of sphingosine (SPH), sphingosine 1-
phosphate (S1P), FTY720 and FTY720-phosphate (FTY720P) on SPC-induced K8 phosphorylation and reorganization
using Western blot and confocal microscopy, and also evaluated the elasticity of PANC-1 cells by atomic force
microscopy. FTY720, FTY720P, SPH, and S1P concentration-dependently inhibited SPC-evoked phosphorylation and
reorganization of K8, and migration of PANC-1 cells. SPC triggered reduction and narrow distribution of elastic
constant K and conversely, FTY720 blocked them. A common upstream regulator of JNK and ERK, protein
phosphatase 2A (PP2A) expression was reduced by SPC, but was restored by FTY720 and FTY72P. Butyryl forskolin,
a PP2A activator, suppressed SPC-induced K8 phosphorylation and okadaic acid, a PP2A inhibitor, induced K8
phosphorylation. Gene silencing of PP2A also led to K8 phosphorylation, reorganization and migration. We also
investigated the involvement of GPR12, a high-affinity SPC receptor, in SPC-evoked keratin phosphorylation and
reorganization. GPR12 siRNA suppressed the SPC-triggered phosphorylation and reorganization of K8. GPR12
overexpression stimulated keratin phosphorylation and reorganization even without SPC. FTY720 and FTY720P
suppressed the GPR12-induced phosphorylation and reorganization of K8. The collective data indicates that FTY720
and FTY720P suppress SPC-induced phosphorylation and reorganization of K8 in PANC-1 cells by restoring the
expression of PP2A via GPR12. These findings might be helpful in the development of compounds that modulate the
viscoelasticity of metastatic cancer cells and various SPC actions.
~1 Citing
301. Obesity-induced p53 activation in insulin-dependent and independent tissues is inhibited by beta-
adrenergic agonist in diet-induced obese rats
By Zand Hamid; Homayounfar Reza; Cheraghpour Makan; Pourvali Katayoun; Jeddi-Tehrani Mahmood; Ghorbani
Arman; Soltani Sama Reza
From Life sciences (2016), 147103-9, Language: English, Database: MEDLINE
AIMS: The purpose of this study was to assay the role of beta-adrenergic receptor signaling in the regulation of
obesity-induced p53 in high fat feeding obese rats. MAIN METHODS: The role of beta-adrenergic receptor/cyclic
AMP in the regulation of p53 and its downstream mediators was evaluated by western blot and real-time quantitative
RT-PCR among diet induced rats. KEY FINDINGS: Beta-adrenergic receptor agonist, isoproterenol, and an adenylate
cyclase activator, forskolin, at a single dose significantly reduced insulin resistance consistent with a decrease in total
and phospho-p53 levels in insulin and non-insulin metabolic target tissues. The decrease of p53 signaling was
consistent with the elevation of AKT and subsequent activation. Obese rats exposed to fasting also exhibited
improvement in insulin action despite a slight effect on p53 level. SIGNIFICANCE: Results of the present study
obviously showed that beta-adrenergic receptor agonist/cAMP prevented obesity-induced p53 activation. Although this
effect in metabolic insulin target tissues tempted us to consider them as insulin sensitizers in obesity-related diabetes,
p53 inhibition in non-insulin target tissues warned about the impairment of anti-cancer mechanisms in obese subjects.
~2 Citings
302. cAMP signaling increases histone deacetylase 8 expression by inhibiting JNK-dependent degradation via
autophagy and the proteasome system in H1299 lung cancer cells
By Park Ji-Yeon; Juhnn Yong-Sung
From Biochemical and biophysical research communications (2016), 470(2), 336-342, Language: English, Database:
MEDLINE
This study aimed to investigate the roles of autophagy and the ubiquitin-proteasome system in the degradation of
histone deacetylase 8 (HDAC8) and to clarify the mechanism by which cAMP signaling regulates this degradation.
cAMP signaling was activated by treating H1299 non-small cell lung cancer cells with isoproterenol or forskolin/3-
isobutyl-1-methylxanthine, and HDAC8 expression was assessed by western blot analysis. The inhibition of autophagy
and ubiquitin-proteasome-dependent degradation increased HDAC8 expression. cAMP signaling inhibited JNK
activation, which decreased the phosphorylation of Bcl-2, thereby reducing autophagy, and the phosphorylation of Itch,
thereby reducing ubiquitination. These results suggest that the HDAC8 protein is degraded via autophagy and the
ubiquitin-proteasome system and that cAMP signaling increases HDAC8 protein levels by reducing JNK-mediated
autophagy and ubiquitin-proteasome-dependent degradation of the HDAC8 protein in H1299 lung cancer cells.
~1 Citing
303. High-Throughput Screening of Chemical Effects on Steroidogenesis Using H295R Human Adrenocortical
Carcinoma Cells
By Karmaus Agnes L; Filer Dayne L; Martin Matthew T; Toole Colleen M; Lewis Kenneth C
From Toxicological sciences : an official journal of the Society of Toxicology (2016), 150(2), 323-32, Language:
English, Database: MEDLINE
Disruption of steroidogenesis by environmental chemicals can result in altered hormone levels causing adverse
reproductive and developmental effects. A high-throughput assay using H295R human adrenocortical carcinoma cells
was used to evaluate the effect of 2060 chemical samples on steroidogenesis via high-performance liquid
chromatography followed by tandem mass spectrometry quantification of 10 steroid hormones, including progestagens,
glucocorticoids, androgens, and estrogens. The study employed a 3 stage screening strategy. The first stage
established the maximum tolerated concentration (MTC; ≥ 70% viability) per sample. The second stage quantified
changes in hormone levels at the MTC whereas the third stage performed concentration-response (CR) on a subset of
samples. At all stages, cells were prestimulated with 10 µM forskolin for 48 h to induce steroidogenesis followed by
chemical treatment for 48 h. Of the 2060 chemical samples evaluated, 524 samples were selected for 6-point CR
screening, based in part on significantly altering at least 4 hormones at the MTC. CR screening identified 232
chemical samples with concentration-dependent effects on 17β-estradiol and/or testosterone, with 411 chemical
samples showing an effect on at least one hormone across the steroidogenesis pathway. Clustering of the
concentration-dependent chemical-mediated steroid hormone effects grouped chemical samples into 5 distinct profiles
generally representing putative mechanisms of action, including CYP17A1 and HSD3B inhibition. A distinct pattern
was observed between imidazole and triazole fungicides suggesting potentially distinct mechanisms of action. From a
chemical testing and prioritization perspective, this assay platform provides a robust model for high-throughput
screening of chemicals for effects on steroidogenesis.
~0 Citings

304. Escherichia coli Heat-Stable Enterotoxin Mediates Na+/H+ Exchanger 4 Inhibition Involving cAMP in T84
Human Intestinal Epithelial Cells
By Beltran Ana R; Cornejo Marcelo; Norambuena Katrina; Ramirez Marco A; Beltran Ana R; Carraro-Lacroix Luciene
R; Bezerra Camila N A; Malnic Gerhard; Toledo Fernando; Araos Joaquin; et al
From PloS one (2015), 10(12), e0146042, Language: English, Database: MEDLINE
The enterotoxigenic Escherichia coli strains lead to diarrhoea in humans due to heat-labile and heat-stable (STa)
enterotoxins. STa increases Cl-release in intestinal cells, including the human colonic carcinoma T84 cell line,
involving increased cGMP and membrane alkalization due to reduced Na+/H+ exchangers (NHEs) activity. Since
NHEs modulate intracellular pH (pHi), and NHE1, NHE2, and NHE4 are expressed in T84 cells, we characterized the
STa role as modulator of these exchangers. pHi was assayed by the NH4Cl pulse technique and measured by
fluorescence microscopy in BCECF-preloaded cells. pHi recovery rate (dpHi/dt) was determined in the absence or
presence of 0.25 µmol/L STa (30 minutes), 25 µmol/L HOE-694 (concentration inhibiting NHE1 and NHE2), 500 µmol/L
sodium nitroprusside (SNP, spontaneous nitric oxide donor), 100 µmol/L dibutyryl cyclic GMP (db-cGMP), 100 nmol/L
H89 (protein kinase A inhibitor), or 10 µmol/L forskolin (adenylyl cyclase activator). cGMP and cAMP were measured in
cell extracts by radioimmunoassay, and buffering capacity (ssi) and H+ efflux (JH+) was determined. NHE4 protein
abundance was determined by western blotting. STa and HOE-694 caused comparable reduction in dpHi/dt and JH+
(~63%), without altering basal pHi (range 7.144-7.172). STa did not alter ssi value in a range of 1.6 pHi units. The
dpHi/dt and JH+ was almost abolished (~94% inhibition) by STa + HOE-694. STa effect was unaltered by db-cGMP or
SNP. However, STa and forskolin increased cAMP level. STa-decreased dpHi/dt and JH+ was mimicked by forskolin,
and STa + HOE-694 effect was abolished by H89. Thus, incubation of T84 cells with STa results in reduced NHE4
activity leading to a lower capacity of pHi recovery requiring cAMP, but not cGMP. STa effect results in a causal
phenomenon (STa/increased cAMP/increased PKA activity/reduced NHE4 activity) ending with intracellular
acidification that could have consequences in the gastrointestinal cells function promoting human diarrhoea.
~1 Citing
305. Modulation of glucocorticoid, mineralocorticoid and androgen production in H295 cells by Trimesemine®, a
mesembrine-rich Sceletium extract
By Swart A C; Smith C
From Journal of ethnopharmacology (2016), 17735-45, Language: English, Database: MEDLINE
ETHNOPHARMACOLOGICAL RELEVANCE: Stress-related illnesses rate among the most prevalent non-fatal
diseases globally. With the global trend for consumer bias towards natural medicine, the Sceletium plant has become
more prominent in the field of natural products. Although potentially useful effects of Sceletium tortuosum on the
central nervous system have been reported, limited data is available on effects of the plant in the peripheral
compartment. AIM OF THE STUDY: The current study aimed to elucidate the effect(s) of a Sceletium extract (TRI)
rich in mesembrine (1% of plant extract w/w), on adrenal steroid biosynthesis. MATERIALS AND METHODS:
Steroidogenesis was assessed basally and in response to stimuli (forskolin, angiotensin II, KCl), in human
adrenocortical carcinoma cells (H295R). Steroid hormone levels were assessed using UPLC-MS/MS. UPLC-MS
analyses of TRI identified major alkaloids ∆7-mesembrenone, mesembrenone and mesembrine. RESULTS: Highest
dose TRI treatment (1 mg/ml, 34.5 µM mesembrine) increased pregnenolone and decreased 16-hydroxyprogesterone
levels (both P<0.00001) in forskolin-stimulated conditions only, suggesting CYP17 enzyme inhibition. This led to
significant inhibition of forskolin-associated increases in cortisol levels at the highest dose (P<0.001) and basal cortisol
levels across all doses (P<0.0001). Independently of forskolin, TRI inhibited androstenedione and testosterone
production across all doses (both P<0.00001), suggesting inhibition of 3βHSD and 17βHSD respectively. TRI
decreased both the angiotensin II- (P<0.05) and forskolin-induced (P<0.0001) increases in aldosterone production.
CONCLUSIONS: Our data suggest potentially beneficial effects of TRI in the context of stress and hypertension.
These should be further investigated in a whole organism model, while the effects on the androgenic pathway should
also be further elucidated.
~0 Citings
306. Role of extracellular calcitonin gene-related peptide in spinal cord mechanisms of cancer-induced bone
pain
By Hansen Rikke R; Vacca Valentina; Pitcher Thomas; Clark Anna K; Malcangio Marzia
From Pain (2016), 157(3), 666-76, Language: English, Database: MEDLINE
Severe pain is a common and debilitating complication of metastatic bone cancer. Current analgesics provide
insufficient pain relief and often lead to significant adverse effects. In models of cancer-induced bone pain,
pathological sprouting of sensory fibers at the tumor-bone interface occurs concomitantly with reactive astrocytosis in
the dorsal horn of the spinal cord. We observed that calcitonin gene-related peptide (CGRP)-fiber sprouting in the
bone was associated with an increase in CGRP content in sensory neuron cell bodies in the dorsal root ganglia (DRG)
and increased basal and activity-evoked release of CGRP from their central terminals in the dorsal horn. Intrathecal
administration of a peptide antagonist (α-CGRP8-37) attenuated referred allodynia in the hind paw ipsilateral to bone
cancer. CGRP receptor components (CLR and RAMP1) were up-regulated in dorsal horn neurons and expressed by
reactive astrocytes. In primary cultures of astrocytes, CGRP incubation led to a concentration-dependent increase of
forskolin-induced cAMP production, which was attenuated by pretreatment with CGRP8-37. Furthermore, CGRP
induced ATP release in astrocytes, which was inhibited by CGRP8-37. We suggest that the peripheral increase in
CGRP content observed in cancer-induced bone pain is mirrored by a central increase in the extracellular levels of
CGRP. This increase in CGRP not only may facilitate glutamate-driven neuronal nociceptive signaling but also act on
astrocytic CGRP receptors and lead to release of ATP.
~1 Citing
307. Low-Dose Endothelial Monocyte-Activating Polypeptide-II Induces Blood-Tumor Barrier Opening Via the
cAMP/PKA/Rac1 Pathway
By Li Zhen; Liu Yun-hui; Liu Jing; Teng Hao; Xi Zhuo; Yao Yi-long; Liu Xiao-bai; Xue Yi-xue; Xue Yi-xue
From Journal of molecular neuroscience : MN (2016), 58(2), 153-61, Language: English, Database: MEDLINE
Previous studies have demonstrated that low-dose endothelial monocyte-activating polypeptide-II (EMAP-II) induces
blood-tumor barrier (BTB) hyperpermeability via both paracellular and transcellular pathways. In a recent study, we
revealed that cyclic adenosine monophosphate (cAMP)/protein kinase A (PKA)-dependent signaling pathway is
involved in EMAP-II-induced BTB hyperpermeability. This study further investigated the exact mechanisms through
which the cAMP/PKA-dependent signaling pathway affects EMAP-II-induced BTB hyperpermeability. In an in vitro
BTB model, low-dose EMAP-II (0.05 nM) induced a significant decrease in Rac1 activity in rat brain microvascular
endothelial cells (RBMECs). Pretreatment with forskolin to elevate intracellular cAMP concentration completely
blocked EMAP-II-induced inactivation of Rac1. Besides, pretreatment with 6Bnz-cAMP to activate PKA partially
attenuated EMAP-II-induced Rac1 inactivation. Moreover, 6Bnz-cAMP pretreatment significantly diminished EMAP-II-
induced changes in BTB permeability, myosin light chain (MLC) phosphorylation, expression and distribution of ZO-1,
and actin cytoskeleton arrangement in RBMECs. These effects of 6Bnz-cAMP were completely blocked in the
presence of NSC-23766 (the specific inhibitor of Rac1). In conclusion, this study demonstrates that low-dose EMAP-II
induces BTB hyperpermeability via the cAMP/PKA/Rac1 signaling pathway.
~1 Citing
308. Atypical signaling of metabotropic glutamate receptor 1 in human melanoma cells

By Gelb Tara; Pshenichkin Sergey; Hathaway Hannah A; Grajkowska Ewa; Dalley Carrie Bowman; Wolfe Barry B;
Wroblewski Jarda T
From Biochemical pharmacology (2015), 98(1), 182-9, Language: English, Database: MEDLINE
The metabotropic glutamate 1 (mGlu1) receptor has emerged as a novel target for the treatment of metastatic
melanoma and various other cancers. Our laboratory has demonstrated that a selective, non-competitive mGlu1
receptor antagonist slows human melanoma growth in vitro and in vivo. In this study, we sought to determine if the
activation of a canonical G protein-dependent signal transduction cascade, which is often used as an output of mGlu1
receptor activity in neuronal cells, correlated with mGlu1 receptor-mediated melanoma cell viability. Glutamate, the
endogenous ligand of mGlu1 receptors, significantly increased melanoma cell viability, but did not stimulate
phosphoinositide (PI) hydrolysis in several human melanoma cell lines. In contrast, melanoma cell viability was not
increased by quisqualate, a highly potent mGlu1 receptor agonist, or DHPG, a selective group I mGlu receptor agonist.
Similarly to glutamate, quisqualate also failed to stimulate PI hydrolysis in mGlu1 receptor-expressing melanoma cells.
These results suggest that the canonical G protein-dependent signal transduction cascade is not coupled to mGlu1
receptors in all human melanoma cells. On the other hand, dynamin inhibition selectively decreased viability of mGlu1
receptor-expressing melanoma cells, suggesting that a mechanism requiring internalization may control melanoma cell
viability. Taken together, these data demonstrate that the approaches commonly used to study mGlu1 receptor
function and signaling in other systems may be inappropriate for studying mGlu1 receptor-mediated melanoma cell
viability.
~1 Citing
309. Posttranscriptional regulation of T-type Ca(2+) channel expression by interleukin-6 in prostate cancer cells
By Weaver Erika M; Hearne Jennifer L; Zamora Francis J; Martin-Caraballo Miguel
From Cytokine (2015), 76(2), 309-20, Language: English, Database: MEDLINE
BACKGROUND: At early stages, the growth of prostate cancers is androgen dependent. At later stages, however, the
growth of prostate cancers becomes androgen independent, which leads to an increase in mortality. The switch to an
androgen-refractory state is associated with neuroendocrine differentiation (NED) of prostate cancer cells. Several
factors including interleukin-6 (IL-6) and increased cAMP production promote NED of prostate cancer cells. In this
work we investigated whether IL-6 evoked NED of LNCaP cells results in a significant change in T-type Ca(2+) channel
expression in comparison to non-stimulated LNCaP cells. METHODS: T-type Ca(2+) channel subunit Cav3.2
expression was studied using PCR analysis, western blot and whole cell recordings. Tubulin IIIβ expression and
neurite-like morphology was assessed to investigate the role of T-type Ca(2+) channels in the differentiation of prostate
cancer cells. RESULTS: Treatment of LNCaP cells with IL-6 for 4days evokes considerable morphological and
biochemical changes consistent with NED. Transcripts of the T-type Ca(2+) channel subunit Cav3.2 but not Cav3.1 or
Cav3.3 are detected in IL-6 stimulated cells. Real time PCR analysis of IL-6 stimulated cells indicates no significant
change in Cav3.2 mRNA expression in comparison to non-stimulated cells. LNCaP cells stimulated with IL-6 show a
threefold increase in T-type Ca(2+) channel subunit Cav3.2 protein expression, suggesting that channel expression is
upregulated by a posttranscriptional mechanism. Electrophysiological recordings reveal that increased Cav3.2 protein
expression following IL-6 stimulation of LNCaP cells does not result in increased expression of functional channels in
the membrane. Functional expression of Cav3.2 channels in LNCaP cells is facilitated by co-stimulation with IL-6 and
the cAMP-stimulating agent, forskolin (FSK). Inhibition of T-type Ca(2+) channel activity in IL-6 stimulated LNCaP cells
prevents the development of morphological characteristics consistent with NED. CONCLUSIONS: These results
indicate that the functional expression of T-type Ca(2+) channels is regulated by the interplay of multiple factors in
LNCaP cells.
~2 Citings
310. Activation of the Tumor Suppressor PP2A Emerges as a Potential Therapeutic Strategy for Treating
Prostate Cancer
By Cristobal Ion; Torrejon Blanca; Garcia-Foncillas Jesus; Gonzalez-Alonso Paula; Daoud Lina; Solano Esther; Manso
Rebeca; Madoz-Gurpide Juan; Rojo Federico
From Marine drugs (2015), 13(6), 3276-86, Language: English, Database: MEDLINE
Protein phosphatase 2A (PP2A) is a tumor suppressor complex that has recently been reported as a novel and highly
relevant molecular target in prostate cancer (PCa). However, its potential therapeutic value remains to be fully
clarified. We treated PC-3 and LNCaP cell lines with the PP2A activators forskolin and FTY720 alone or combined
with the PP2A inhibitor okadaic acid. We examined PP2A activity, cell growth, prostasphere formation, levels of PP2A
phosphorylation, CIP2A and SET expression, and AKT and ERK activation. Interestingly, both forskolin and FTY720
dephosphorylated and activated PP2A, impairing proliferation and prostasphere formation and inducing changes in
AKT and ERK phosphorylation. Moreover, FTY720 led to reduced CIP2A levels. Treatment with okadaic acid
impaired PP2A activation thus demonstrating the antitumoral PP2A-dependent mechanism of action of both forskolin
and FTY720. Levels of PP2A phosphorylation together with SET and CIP2A protein expression were studied in 24
PCa patients and both were associated with high Gleason scores and presence of metastatic disease. Altogether, our
results suggest that PP2A inhibition could be involved in PCa progression, and the use of PP2A-activating drugs might
represent a novel alternative therapeutic strategy for treating PCa patients.
~2 Citings
311. Inhibition of breast cancer cell migration by activation of cAMP signaling

By Dong Hongli; Claffey Kevin P; Epstein Paul M; Brocke Stefan
From Breast cancer research and treatment (2015), 152(1), 17-28, Language: English, Database: MEDLINE
Almost all deaths from breast cancer arise from metastasis of the transformed cells to other sites in the body. Hence,
uncovering a means of inhibiting breast cancer cell migration would provide a significant advance in the treatment of
this disease. Stimulation of the cAMP signaling pathway has been shown to inhibit migration and motility of a number
of cell types. A very effective way of selectively stimulating cAMP signaling is through inhibition of cyclic nucleotide
phosphodiesterases (PDEs). Therefore, we examined full expression profiles of all known PDE genes at the mRNA
and protein levels in four human breast cancer cell lines and eight patients' breast cancer tissues. By these analyses,
expression of almost all PDE genes was seen in both cell lines and tissues. In the cell lines, appreciable expression
was seen for PDEs 1C, 2A, 3B, 4A, 4B, 4D, 5A, 6B, 6C, 7A, 7B, 8A, 9A, 10A, and 11A. In patients' tissues,
appreciable expression was seen for PDEs 1A, 3B, 4A, 4B, 4C, 4D, 5A, 6B, 6C, 7A, 7B, 8A, 8B, and 9A. PDE8A
mRNA in particular is prominently expressed in all cell lines and patients' tissue samples examined. We show here
that stimulation of cAMP signaling with cAMP analogs, forskolin, and PDE inhibitors, including selective inhibitors of
PDE3, PDE4, PDE7, and PDE8, inhibit aggressive triple negative MDA-MB-231 breast cancer cell migration. Under
the same conditions, these agents had little effect on breast cancer cell proliferation. This study demonstrates that
PDE inhibitors inhibit breast cancer cell migration, and thus may be valuable therapeutic targets for inhibition of breast
cancer metastasis. Since PDE8A is expressed in all breast cancer samples, and since dipyridamole, which inhibits
PDE8, and PF-04957325, a selective PDE8 inhibitor, both inhibit migration, it suggests that PDE8A may be a valuable
novel target for treatment of this disease.
~4 Citings
312. DuOx2 Promoter Regulation by Hormones, Transcriptional Factors and the Coactivator TAZ
By Cardoso-Weide L C; Cardoso-Penha R C; Carvalho D P; Costa M W; Ferreira A C F; Santisteban P S
From European thyroid journal (2015), 4(1), 6-13, Language: English, Database: MEDLINE
The production of H2O2, which is essential to thyroid hormone synthesis, involves two NADPH oxidases: dual
oxidases 1 and 2 (DuOx1 and DuOx2). A functional study with human DuOx genes and their 5'-flanking regions
showed that DuOx1 and -2 promoters are different from thyroid-specific gene promoters. Furthermore, their
transcriptional activities are not restricted to thyroid cells. While regulation of Tg (thyroglobulin) and TPO
(thyroperoxidase) expression have been extensively studied, DuOx2 promoter regulation by hormones and
transcriptional factors need to be more explored. Herein we investigated the role of TSH, insulin and insulin-like
growth factor 1 (IGF-1), as well as the cAMP effect on DuOx2 promoter (ptx41) activity in transfected rat thyroid cell
lines (PCCL3). We also assessed DuOx2 promoter activity in the presence of transcriptional factors crucial to thyroid
development such as TTF-1 (thyroid transcription factor 1), PAX8, CREB, DREAM, Nkx2.5 and the coactivator TAZ in
HeLa and HEK 293T-transfected cells. Our results show that TSH and forskolin, which increase cAMP in thyroid cells,
stimulated DuOx2 promoter activity. IGF-1 led to pronounced stimulation, while insulin induction was not statistically
different from DuOx2 promoter basal activity. All transcriptional factors selected for this work and coactivator TAZ,
except DREAM, stimulated DuOx2 promoter activity. Moreover, Nkx2.5 and TAZ synergistically increased DuOx2
promoter activity. In conclusion, we show that DuOx2 expression is regulated by hormones and transcription factors
involved in thyroid organogenesis and carcinogenesis, reinforcing the importance of the control of H2O2 generation in
the thyroid.
~1 Citing
313. Perfluorooctyl Iodide Stimulates Steroidogenesis in H295R Cells via a Cyclic Adenosine Monophosphate
Signaling Pathway
By Wang Chang; Wang Chang; Ruan Ting; Liu Jiyan; He Bin; Zhou Qunfang; Jiang Guibin
From Chemical research in toxicology (2015), 28(5), 848-54, Language: English, Database: MEDLINE
Perfluorinated iodine alkanes (PFIs) are used widely in the organic fluorine industry. Increased production of PFIs has
caused environmental health concerns. To evaluate the potential endocrine-disrupting effect of PFIs, we investigated
the effects of perfluorooctyl iodide (PFOI) on steroidogenesis in human adrenocortical carcinoma cells (H295R).
Levels of aldosterone, cortisol, 17β-estradiol, and testosterone were measured in H295R culture medium upon
treatment with perfluorooctanoic acid (PFOA) and PFIs. Expression of 10 steroidogenic genes (StAR, HMGR,
CYP11A1, 3βHSD2, 17βHSD, CYP17, CYP21, CYP11B1, CYP11B2, and CYP19) was measured by real-time
polymerase chain reaction. Levels of cyclic adenosine monophosphate (cAMP) and adenylate cyclase (AC) activity
were measured to understand the underlying mechanism of steroidogenic perturbations. Levels of production of
aldosterone, cortisol, and 17β-estradiol were elevated significantly, and the level of testosterone generation decreased
upon treatment with 100 µM PFOI. Similar to the effect induced by forskolin (AC activator), expression of all 10 genes
involved in the synthesis of steroid hormones was upregulated significantly upon exposure to 100 µM PFOI. PFOA
had no effect on steroid hormone production or steroidogenic gene expression even though it is highly structurally
similar with PFOI. Therefore, the terminal -CF2I group in PFOI could be a critical factor for mediation of
steroidogenesis. PFOI increased AC activity and cAMP levels in H295R cells, which implied an underlying mechanism
for the disturbance of steroidogenesis. These data suggest that PFOI may act as an AC activator, thereby stimulating
steroidogenesis by activating a cAMP signaling pathway.
~0 Citings
314. Advances in the analytical methodologies: Profiling steroids in familiar pathways-challenging dogmas
By Bloem Liezl M; Storbeck Karl-Heinz; Swart Pieter; du Toit Therina; Schloms Lindie; Swart Amanda C
From The Journal of steroid biochemistry and molecular biology (2015), 15380-92, Language: English, Database:
MEDLINE
The comprehensive evaluation of the adrenal steroidogenic pathway, given its complexity, requires methodology
beyond the standard techniques currently employed. Advances in LC-MS/MS, coupled with in vitro cell models that
produce all the steroid metabolites of the mineralo-, glucocorticoid and androgen arms, present a powerful approach
for the comprehensive evaluation of adrenal steroidogenesis in response to compounds of interest including bioactives,
drug treatments and endocrine disrupting compounds. UHPLC-MS/MS analysis of steroid panels in forskolin,
angiotensin II and K(+) stimulated H295R cells provides a snapshot of their effect on intermediates and end products
of adrenal steroidogenesis. The impact of full steroid panel evaluations by LC- and GC-MS/MS extends to clinical
profiling with the characterization of normal pediatric steroid reference ranges in sexual development and of disease-
specific profiles improving diagnosis and sub classification. Comprehensive analyses of steroid profiles may potentially
improve patient outcomes together with the application of treatments specifically suited to clinical subgroups. LC-
MS/MS and GC-MS/MS applications in the analyses of comprehensive steroid panels are demonstrated in clinical
conditions such as congenital adrenal hyperplasia in newborns requiring accurate diagnoses and in predicting
metabolic risk in polycystic ovary syndrome patients. Most notable perhaps is the impact of LC-MS/MS evaluations on
our understanding of the basic biochemistry of steroidogenesis with the detection of the long forgotten adrenal steroid,
11β-hydroxyandrostenedione, at significant levels. The characterization of its metabolism to androgen receptor ligands
in the LNCaP prostate cancel cell model, specifically within the context of recurring prostate cancer, lends new
perspectives to old dogmas. We demonstrate that UHPLC-MS/MS has enabled the analyses of novel metabolites of
the enzymes, SRD5A, 11βHSD and 17βHSD, in LNCaP cells. Undoubtedly, the continuous advances in the analytical
methodologies used for steroid profiling and quantification will give impetus to the unraveling of the remaining enigmas,
old and new, of both hormone biosynthesis and metabolism.
~2 Citings
315. The 4'-hydroxyl group of resveratrol is functionally important for direct activation of PPARα
By Takizawa Yoshie; Nakata Rieko; Inoue Hiroyasu; Fukuhara Kiyoshi; Yamashita Hiroshi; Kubodera Hideo
Long-term moderate consumption of red wine is associated with a reduced risk of developing lifestyle-related diseases
such as cardiovascular disease and cancer. Therefore, resveratrol, a constituent of grapes and various other plants,
has attracted substantial interest. This study focused on one molecular target of resveratrol, the peroxisome
proliferator activated receptor α (PPARα). Our previous study in mice showed that resveratrol-mediated protection of
the brain against stroke requires activation of PPARα; however, the molecular mechanisms involved in this process
remain unknown. Here, we evaluated the chemical basis of the resveratrol-mediated activation of PPARα by
performing a docking mode simulation and examining the structure-activity relationships of various polyphenols. The
results of experiments using the crystal structure of the PPARα ligand-binding domain and an analysis of the activation
of PPARα by a resveratrol analog 4-phenylazophenol (4-PAP) in vivo indicate that the 4'-hydroxyl group of resveratrol
is critical for the direct activation of PPARα. Activation of PPARα by 5 µM resveratrol was enhanced by rolipram, an
inhibitor of phosphodiesterase (PDE) and forskolin, an activator of adenylate cyclase. We also found that resveratrol
has a higher PDE inhibitory activity (IC50 = 19 µM) than resveratrol analogs trans-4-hydroxystilbene and 4-PAP (IC50
= 27-28 µM), both of which has only 4'-hydroxyl group, indicating that this 4'-hydroxyl group of resveratrol is not
sufficient for the inhibition of PDE. This result is consistent with that 10 µM resveratrol has a higher agonistic activity of
PPARα than these analogs, suggesting that there is a feedforward activation loop of PPARα by resveratrol, which may
be involved in the long-term effects of resveratrol in vivo.
~4 Citings
316. Cyclic AMP signaling reduces sirtuin 6 expression in non-small cell lung cancer cells by promoting
ubiquitin-proteasomal degradation via inhibition of the Raf-MEK-ERK (Raf/mitogen-activated extracellular signal-
regulated kinase/extracellular signal-regulated kinase) pathway
By Kim Eui-Jun; Juhnn Yong-Sung
From The Journal of biological chemistry (2015), 290(15), 9604-13, Language: English, Database: MEDLINE
The cAMP signaling system regulates various cellular functions, including metabolism, gene expression, and death.
Sirtuin 6 (SIRT6) removes acetyl groups from histones and regulates genomic stability and cell viability. We
hypothesized that cAMP modulates SIRT6 activity to regulate apoptosis. Therefore, we examined the effects of cAMP
signaling on SIRT6 expression and radiation-induced apoptosis in lung cancer cells. cAMP signaling in H1299 and
A549 human non-small cell lung cancer cells was activated via the expression of constitutively active Gαs plus
treatment with prostaglandin E2 (PGE2), isoproterenol, or forskolin. The expression of sirtuins and signaling molecules
were analyzed by Western blotting. Activation of cAMP signaling reduced SIRT6 protein expression in lung cancer
cells. cAMP signaling increased the ubiquitination of SIRT6 protein and promoted its degradation. Treatment with
MG132 and inhibiting PKA with H89 or with a dominant-negative PKA abolished the cAMP-mediated reduction in
SIRT6 levels. Treatment with PGE2 inhibited c-Raf activation by increasing inhibitory phosphorylation at Ser-259 in a
PKA-dependent manner, thereby inhibiting downstream MEK-ERK signaling. Inhibiting ERK with inhibitors or with
dominant-negative ERKs reduced SIRT6 expression, whereas activation of ERK by constitutively active MEK
abolished the SIRT6-depleting effects of PGE2. cAMP signaling also augmented radiation-induced apoptosis in lung
cancer cells. This effect was abolished by exogenous expression of SIRT6. It is concluded that cAMP signaling
reduces SIRT6 expression by promoting its ubiquitin-proteasome-dependent degradation, a process mediated by the
PKA-dependent inhibition of the Raf-MEK-ERK pathway. Reduced SIRT6 expression mediates the augmentation of
radiation-induced apoptosis by cAMP signaling in lung cancer cells.
~6 Citings
317. Therapeutic Re-Activation of Protein Phosphatase 2A in Acute Myeloid Leukemia

By Ramaswamy Kavitha; Spitzer Barbara; Kentsis Alex
From Frontiers in oncology (2015), 516, Language: English, Database: MEDLINE
Protein phosphatase 2A (PP2A) is a serine/threonine phosphatase that is required for normal cell growth and
development. PP2A is a potent tumor suppressor, which is inactivated in cancer cells as a result of genetic deletions
and mutations. In myeloid leukemias, genes encoding PP2A subunits are generally intact. Instead, PP2A is
functionally inhibited by post-translational modifications of its catalytic C subunit, and interactions with negative
regulators by its regulatory B and scaffold A subunits. Here, we review the molecular mechanisms of genetic and
functional inactivation of PP2A in human cancers, with a particular focus on human acute myeloid leukemias (AML).
By analyzing expression of genes encoding PP2A subunits using transcriptome sequencing, we find that PP2A
dysregulation in AML is characterized by silencing and overexpression of distinct A scaffold and B regulatory subunits,
respectively. We review the mechanisms of functional PP2A activation by drugs such as fingolimod, forskolin, OP449,
and perphenazine. This analysis yields two non-mutually exclusive mechanisms for therapeutic PP2A re-activation: (i)
allosteric activation of the phosphatase activity, and (ii) stabilization of active holo-enzyme assembly and displacement
of negative regulatory factors from A and B subunits. Future studies should allow the development of specific and
potent pharmacologic activators of PP2A, and definition of susceptible disease subsets based on specific mechanisms
of PP2A dysregulation.
~2 Citings
318. PKA/CREB regulates the constitutive promoter activity of the USP22 gene
By Xiong Jianjun; Zhou Xiaoou; Gong Zhen; Wang Ting; Xu Xiaoyuan; Zhang Chao; Liu Jianyun; Li Weidong
From Oncology reports (2015), 33(3), 1505-11, Language: English, Database: MEDLINE
The human ubiquitin-specific processing enzyme 22 (USP22) plays a crucial role in regulating cell cycle processes and
its overexpression has been linked to tumor progression. However, the mechanisms leading to USP22 transcriptional
activation in human cancer cells are still unclear. Previously, we characterized the 5'-flanking sequence of the human
USP22 gene and found a potential CREB/ATF binding site within the basic promoter region. The present study found
that this site was required for constitutive USP22 transcriptional activity in HeLa and HepG2 cells. Chromatin
immunoprecipitation assay confirmed that CREB interacted with this site. siRNA knockdown of CREB decreased
USP22 transcriptional activation and endogenous expression, whereas CREB overexpression did not affect
transcriptional levels. Furthermore, USP22 promoter activity and expression were decreased by inhibiting PKA with H-
89, but were not responsive to forskolin induction. All of these results demonstrate that PKA/CREB is involved in the
regulation of constitutive promoter activity of the USP22 gene.
~1 Citing
319. MicroRNA-124 regulates glucocorticoid sensitivity by targeting phosphodiesterase 4B in diffuse large B cell
lymphoma
By Kim Jinyoung; Jeong Dawoom; Nam Jehyun; Aung Thazin Nwe; Gim Jeong-An; Park Keon Uk; Kim Sang-Woo
From Gene (2015), 558(1), 173-80, Language: English, Database: MEDLINE
Glucocorticoids (GCs) are chemotherapeutic drugs commonly used to treat hematological malignancies. However, a
significant fraction of patients develop resistance to GCs during treatment. A better insight into how GC resistance
develops is therefore needed. It was previously shown that cyclic AMP (cAMP) induces sensitivity to GCs by inhibiting
the AKT/mTOR/MCL1 signaling, while high levels of phosphodiesterase 4B (PDE4B) reverse the effect of cAMP on GC
responses in B-cell lymphoma. Here, we show that miR-124 influences GC-induced apoptosis by directly targeting
PDE4B. Stable expression of miR-124 in diffuse large B cell lymphoma (DLBCL) cell lines diminished PDE4B
expression. This was associated with increased cAMP levels, inhibition of the AKT/mTOR/MCL1 survival pathway,
upregulation of GRα expression, and improved sensitivity to GCs in the presence of forskolin, an activator of adenylyl
cyclase. Interestingly, miR-124 did not affect GC sensitivity in the absence of forskolin, indicating that the effect of this
miRNA is accomplished via downregulation of PDE4B expression. Further, restoration of PDE4B expression in miR-
124 cells rescued the phenotypic effect of this miRNA, demonstrating the critical role of PDE4B in miR-124-mediated
regulation of the GC response. Our study supports the notion that miR-124 could be an attractive therapeutic target for
overcoming GC resistance in DLBCL.
~3 Citings
320. Salt-inducible kinase 2 regulates mitotic progression and transcription in prostate cancer
By Bon Helene; Wadhwa Karan; Ramos-Montoya Antonio; Ross-Adams Helen; Neal David E; Mills Ian G; Schreiner
Alexander; Osborne Michelle; Carroll Thomas; Visser Matthieu; et al
From Molecular cancer research : MCR (2015), 13(4), 620-635, Language: English, Database: MEDLINE
UNLABELLED: Salt-inducible kinase 2 (SIK2) is a multifunctional kinase of the AMPK family that plays a role in
CREB1-mediated gene transcription and was recently reported to have therapeutic potential in ovarian cancer. The
expression of this kinase was investigated in prostate cancer clinical specimens. Interestingly, auto-antibodies against
SIK2 were increased in the plasma of patients with aggressive disease. Examination of SIK2 in prostate cancer cells
found that it functions both as a positive regulator of cell-cycle progression and a negative regulator of CREB1 activity.
Knockdown of SIK2 inhibited cell growth, delayed cell-cycle progression, induced cell death, and enhanced CREB1
activity. Expression of a kinase-dead mutant of SIK2 also inhibited cell growth, induced cell death, and enhanced
CREB1 activity. Treatment with a small-molecule SIK2 inhibitor (ARN-3236), currently in preclinical development, also
led to enhanced CREB1 activity in a dose- and time-dependent manner. Because CREB1 is a transcription factor and
proto-oncogene, it was posited that the effects of SIK2 on cell proliferation and viability might be mediated by changes
in gene expression. To test this, gene expression array profiling was performed and while SIK2 knockdown or
overexpression of the kinase-dead mutant affected established CREB1 target genes; the overlap with transcripts
regulated by forskolin (FSK), the adenylate cyclase/CREB1 pathway activator, was incomplete. IMPLICATIONS: This
study demonstrates that targeting SIK2 genetically or therapeutically will have pleiotropic effects on cell-cycle
progression and transcription factor activation, which should be accounted for when characterizing SIK2 inhibitors.
~5 Citings
321. Role of LKB1-CRTC1 on glycosylated COX-2 and response to COX-2 inhibition in lung cancer
By Cao Chunxia; Gao Ruli; Zhang Min; Amelio Antonio L; Fallahi Mohammad; Chen Zirong; Gu Yumei; Hu Chengbin;
Welsh Eric A; Engel Brienne E; et al
From Journal of the National Cancer Institute (2015), 107(1), 358, Language: English, Database: MEDLINE
BACKGROUND: Cyclooxygenase-2 (COX-2) directs the synthesis of prostaglandins including PGE-2 linking
inflammation with mitogenic signaling. COX-2 is also an anticancer target, however, treatment strategies have been
limited by unreliable expression assays and by inconsistent tumor responses to COX-2 inhibition. METHODS: We
analyzed the TCGA and Director's Challenge lung cancer datasets (n = 188) and also generated an LKB1-null lung
cancer gene signature (n = 53) to search the Broad Institute/Connectivity-MAP (C-MAP) dataset. We performed ChIP
analyses, real-time polymerase chain reaction, immunoblotting, and drug testing of tumor cell lines (n = 8) and primary
lung adenocarcinoma surgical resections (n = 13). RESULTS: We show that COX-2 is a target of the cAMP/CREB
coactivator CRTC1 signaling pathway. In addition, we detected a correlation between LKB1 status, CRTC1 activation,
and presence of glycosylated, but not inactive hypoglycosylated COX-2 in primary lung adenocarcinoma. A search of
the C-MAP drug database discovered that all high-ranking drugs positively associated with the LKB1-null signature are
known CRTC1 activators, including forskolin and six different PGE-2 analogues. Somatic LKB1 mutations are present
in 20.0% of lung adenocarcinomas, and we observed growth inhibition with COX-2 inhibitors in LKB1-null lung cancer
cells with activated CRTC1 as compared with LKB1-wildtype cells (NS-398, P = .002 and Niflumic acid, P = .006; two-
tailed t test). CONCLUSION: CRTC1 activation is a key event that drives the LKB1-null mRNA signature in lung
cancer. We also identified a positive feedback LKB1/CRTC1 signaling loop for COX-2/PGE2 regulation. These data
suggest a role for LKB1 status and glycosylated COX-2 as specific biomarkers that provide a framework for selecting
patients for COX-2 inhibition studies.
~9 Citings
322. Role of cAMP-dependent protein kinase A activity in low-dose endothelial monocyte-activating polypeptide-
II-induced opening of blood-tumor barrier
By Li Zhen; Liu Xiao-bai; Liu Yun-hui; Xue Yi-xue; Wang Ping; Liu Li-bo
From Journal of molecular neuroscience : MN (2015), 56(1), 60-9, Language: English, Database: MEDLINE
Our previous studies demonstrated that low-dose endothelial monocyte-activating polypeptide-II (EMAP-II) can
selectively increase the permeability of blood-tumor barrier (BTB). In addition, low-dose EMAP-II significantly
decreases the cyclic adenosine monophosphate (cAMP) concentration and the protein kinase A (PKA) expression
level in tumor tissues in the rat C6 glioma model. In this study, an in vitro BTB model was used to investigate the
potential role of cAMP/PKA signaling cascade in EMAP-II-induced BTB hyperpermeability. Our data revealed that low-
dose EMAP-II (0.05 nM) induced a significant decrease in total intracellular cAMP concentration and PKA activity in rat
brain microvascular endothelial cells (RBMECs). Pretreatment with forskolin to increase intracellular cAMP nearly
completely blocked the EMAP-II-induced decrease in transendothelial electric resistance and increase in horseradish
peroxidase flux across the BTB. Similar pretreatment completely prevented the EMAP-II-induced changes in
RhoA/Rho kinase activity, expression and distribution of tight junction-associated protein ZO-1, and myosin light chain
phosphorylation, as well as actin cytoskeleton arrangement in RBMECs. Pretreatment with 6Bnz-cAMP to activate
PKA significantly attenuated these EMAP-II-induced alterations in RBMECs. In summary, our present study
demonstrates that the cAMP/PKA signaling cascade works as a crucial signaling pathway in EMAP-II-induced BTB
hyperpermeability.
~0 Citings
323. Synthesis of cyclic 1,9-acetal derivatives of forskolin and their bioactivity evaluation
By Ponnam Devendar; Arigari Niranjan Kumar; Shilpi Singh; Suiab Luqman; Siddiqui Lubna; Dubey Vijaya; Srinivas K
V N S; Jonnala Kotesh Kumar; Alam Sarfaraz; Khan Feroz; et al
From European journal of medicinal chemistry (2014), 87735-44, Language: English, Database: MEDLINE
A new series of 1,9-acetals of forskolin were synthesized by treating with aromatic and aliphatic aldehydes using Ceric
ammonium nitrate as catalyst and evaluated for anticancer and α-glucosidase inhibition activities. Among the
synthesized compounds 2a, 2b and 3a showed potential cytotoxic activity towards human cancer cell lines MCF-7
(Human Breast Adenocarcinoma), MDA-MB (Human Breast Carcinoma), HeLa (Human Cervix Adenocarcinoma),
A498 (Human Kidney Carcinoma), K562 (Human Erythromyeloblastoid leukemia), SH-SY5Y (Human Neuroblastoma),
Hek293 (Human Embryonic Kidney) and WRL68 (Human Hepatic) with IC50 values ranging between 0.95 and 47.96
µg/ml. Osmotic fragility test revealed compound 3a as non-toxic to human erythrocytes at the tested concentrations of
50 and 100 µg/ml. Compounds 1g (IC50 value 0.76 µg/ml) and 1p (IC50 value 0.74 µg/ml) significantly inhibited α-
glucosidase in in vitro system. In silico based docking, ADME and toxicity risk assessment studies also showed
discernible α-glucosidase activity for compounds 1g, 1p compared to standard acarbose.
~1 Citing
324. Glucagon-like peptide-1 stimulates type 3 iodothyronine deiodinase expression in a mouse insulinoma cell
line
By Akiyama Shigeo; Ogiwara Takayuki; Aoki Tomoyuki; Tsunekawa Katsuhiko; Araki Osamu; Murakami Masami
From Life sciences (2014), 115(1-2), 22-8, Language: English, Database: MEDLINE
AIMS: The pathophysiological roles of thyroid hormones in glucose metabolism remain uncertain. Type 3
iodothyronine deiodinase (D3) converts thyroxine (T4) and 3,5,3'-triiodothyronine (T3) to 3,3',5'-triiodothyronine (rT3)
and 3,3'-diiodothyronine (T2), respectively, inactivating thyroid hormones in a cell-specific fashion. In the present
study, we identified D3 expression in MIN6 cells derived from a mouse insulinoma cell line and examined the
mechanisms regulating D3 expression in these cells. MAIN METHODS: We characterized D3 activity using HPLC
analysis, and examined the effect of GLP-1 or exendin-4 on D3 expression and cAMP accumulation in MIN6 cells. We
also measured insulin secretion from MIN6 cells exposed to GLP-1 and T3. KEY FINDINGS: We identified enzyme
activity that catalyzes the conversion of T3 to T2 in MIN6 cells, which showed characteristics compatible with those for
D3. D3 mRNA was identified in these cells using RT-PCR analysis. Forskolin rapidly stimulated D3 mRNA and D3
activity. Glucagon-like peptide-1 (GLP-1) increased D3 expression in a dose-dependent manner, and this effect was
inhibited by the protein kinase A (PKA) inhibitor H-89. Exendin-4, a GLP-1 receptor agonist, also stimulated D3
expression in MIN6 cells. These results suggest that a cAMP-PKA-mediated pathway participates in GLP-1-stimulated
D3 expression in MIN6 cells. Furthermore, GLP-1 stimulated insulin secretion was suppressed by the addition of T3 in
MIN6 cells. SIGNIFICANCE: Our findings indicate that GLP-1 regulates intracellular T3 concentration in pancreatic β
cells via a cAMP-PKA-D3-mediated pathway that may also regulate β-cell function.
~0 Citings

325. Quantitative impedimetric NPY-receptor activation monitoring and signal pathway profiling in living cells
By te Kamp Verena; Lindner Ricco; Jahnke Heinz-Georg; Krinke Dana; Kostelnik Katja B; Beck-Sickinger Annette G;
Robitzki Andrea A
From Biosensors & bioelectronics (2015), 67386-93, Language: English, Database: MEDLINE
Label-free and non-invasive monitoring of receptor activation and identification of the involved signal pathways in living
cells is an ongoing analytic challenge and a great opportunity for biosensoric systems. In this context, we developed
an impedance spectroscopy-based system for the activation monitoring of NPY-receptors in living cells. Using an
optimized interdigital electrode array for sensitive detection of cellular alterations, we were able for the first time to
quantitatively detect the NPY-receptor activation directly without a secondary or enhancer reaction like cAMP-
stimulation by forskolin. More strikingly, we could show that the impedimetric based NPY-receptor activation
monitoring is not restricted to the Y1-receptor but also possible for the Y2- and Y5-receptor. Furthermore, we could
monitor the NPY-receptor activation in different cell lines that natively express NPY-receptors and proof the specificity
of the observed impedimetric effect by agonist/antagonist studies in recombinant NPY-receptor expressing cell lines.
To clarify the nature of the observed impedimetric effect we performed an equivalent circuit analysis as well as
analyzed the role of cell morphology and receptor internalization. Finally, an antagonist based extensive molecular
signal pathway analysis revealed small alterations of the actin cytoskeleton as well as the inhibition of at least L-type
calcium channels as major reasons for the observed NPY-induced impedance increase. Taken together, our novel
impedance spectroscopy based NPY-receptor activation monitoring system offers the opportunity to identify signal
pathways as well as for novel versatile agonist/antagonist screening systems for identification of novel therapeutics in
the field of obesity and cancer.
~0 Citings
326. TD-19, an erlotinib derivative, induces epidermal growth factor receptor wild-type nonsmall-cell lung cancer
apoptosis through CIP2A-mediated pathway
By Chao Ting-Ting; Wang Cheng-Yi; Lai Chih-Cheng; Chen Yen-Lin; Tsai Yi-Ting; Chen Pao-Tzu; Lin Hen-I; Huang
Yuh-Chin T; Shiau Chung-Wai; Yu Chong-Jen; et al
From The Journal of pharmacology and experimental therapeutics (2014), 351(2), 352-8, Language: English,
Database: MEDLINE
Some patients with nonsmall-cell lung cancer (NSCLC) without epidermal growth factor receptor (EGFR) mutations still
respond to gefitinib and erlotinib, suggesting that there may be a mechanism(s) other than the EGFR pathway that
mediates the tumoricidal effects. In the current study, we tested the efficacy of TD-19, a novel compound chemically
modified from erlotinib, which has more potent apoptotic effects than erlotinib in EGFR wild-type NSCLC cell lines. TD-
19 induced significant cell death and apoptosis in H358, H441, H460, and A549 cells, as evidenced by increased
caspase-3 activity and cleavage of procaspase-9 and poly (ADP-ribose) polymerase. The apoptotic effect of TD-19 in
H460 cells, which were resistant to erlotinib, was associated with downregulation of cancerous inhibitor of protein
phosphatase 2A (CIP2A), increased protein phosphatase 2A (PP2A) activity, and decreased AKT phosphorylation, but
minimal effects on EGFR phosphorylation. Overexpression of CIP2A partially protected the H460 cells from TD-19-
induced apoptosis. Okadaic acid, a known PP2A inhibitor, significantly reduced TD-19-induced apoptosis, while
forskolin, which increased PP2A activity, increased the apoptotic effect of TD-19. TD-19 inhibited the growth of H460
xenograft tumors by ∼80%. We conclude that TD-19 exerted tumoricidal effects on NSCLC cells. TD-19 provides
proof that the CIP2A pathway may be a novel target for the treatment of EGFR wild-type NSCLC.
~2 Citings
327. MicroRNA-378a-5p targets cyclin G2 to inhibit fusion and differentiation in BeWo cells
By Nadeem Uzma; Ye Gang; Salem Mohamed; Peng Chun
From Biology of reproduction (2014), 91(3), 76, Language: English, Database: MEDLINE
MicroRNAs are expressed abundantly in the placenta throughout pregnancy. We have previously reported that
microRNA (miR)-378a-5p promoted trophoblast migration and invasion. To further understand the role of miR-378a-5p
during placental development, we investigated whether it may regulate the differentiation of syncytiotrophoblast (STB).
Using a choriocarcinoma cell line, BeWo, we found that miR-378a-5p was down-regulated during forskolin-induced
STB differentiation. Transfection of a miR-378a-5p mimic into BeWo cells decreased the formation of multinucleated
STB, increased E-cadherin, and decreased the expression level of STB marker genes. On the other hand, transfection
of anti-miR-378a-5p resulted in an increase in formation of multinucleated STB and expression of STB marker genes,
as well as the loss of E-cadherin. Bioinformatic analysis revealed that miR-378a-5p has four potential binding sites at
the 3' untranslated region (UTR) of cyclin G2 (CCNG2). Using luciferase reporter assays, we showed that miR-378a-
5p decreased the luciferase activity of reporter constructs that contain CCNG2 3' UTR. In addition, miR-378a-5p
decreased, whereas anti-miR-378a-5p increased, CCNG2 mRNA levels. Overexpression of CCNG2 increased the
expression of syncytin-1 and fusion index and reversed the inhibitory effects of miR-378a-5p. In contrast, silencing of
CCNG2 using siRNA increased E-cadherin and decreased syncytin-1 levels. These findings provide initial evidence
that CCNG2 promotes STB differentiation and suggest that miR-378a-5p exerts an inhibitory role in STB differentiation,
in part, by down-regulating CCNG2 expression, in the BeWo cell model.
~2 Citings
328. Prostaglandin E2 promotes the cell growth and invasive ability of hepatocellular carcinoma cells by
upregulating c-Myc expression via EP4 receptor and the PKA signaling pathway
By Xia Shukai; Ma Juan; Bai Xiaoming; Zhang Hai; Cheng Shanyu; Zhang Min; Zhang Li; Du Mingzhan; Wang Yipin; Li
Hai; et al
From Oncology reports (2014), 32(4), 1521-30, Language: English, Database: MEDLINE
Hepatocellular carcinoma (HCC) represents a major health problem worldwide. Prostaglandin E2 (PGE2), the
predominant product of cyclooxygenase-2, has been implicated in hepatocarcinogenesis. However, the underlying
molecular mechanisms remain to be further elucidated. c-myc, a cellular proto-oncogene, is activated or
overexpressed in many types of human cancer, including HCC. The present study was designed to investigate the
internal relationship and molecular mechanisms between PGE2 and c-Myc in HCC, and to define its role in HCC cell
growth and invasion. Our results showed that PGE2 significantly upregulated c-Myc expression at both the mRNA and
protein levels, and knockdown of c-Myc blocked PGE2-induced HCC cell growth and invasive ability in human HCC
Huh-7 cells. The effect of PGE2 on c-Myc expression was mainly through the EP4 receptor, and EP4 receptor-
mediated c-Myc protein upregulation largely depended on de novo biosynthesis of c-Myc mRNA and its protein. EP4
receptor signaling activated GS/AC and increased the intracellular cAMP level in Huh-7 cells. The adenylate cyclase
(AC) activator forskolin mimicked the effects of the EP4 receptor agonist on c-Myc expression, while the AC inhibitor
SQ22536 reduced EP4 receptor-mediated c-Myc upregulation. These data confirm the involvement of the
GS/AC/cAMP pathway in EP4 receptor-mediated c-Myc upregulation. Moreover, the phosphorylation levels of CREB
protein were markedly elevated by EP4 receptor signaling, and by using specific inhibitor and siRNA interference, we
demonstrated that PKA/CREB was also involved in the EP4 receptor-mediated c-Myc upregulation. In summary, the
present study revealed that PGE2 significantly upregulates c-Myc expression at both mRNA and protein levels through
the EP4R/GS/AC/cAMP/PKA/CREB signaling pathway, thus promoting cell growth and invasion in HCC cells.
Targeting of the PGE2/EP4R/c-Myc pathway may be a new therapeutic strategy to prevent and cure human HCC.
~2 Citings
329. Steroidogenic enzymes, their related transcription factors and nuclear receptors in human sebaceous
glands under normal and pathological conditions
By Azmahani Abdullah; Nakamura Yasuhiro; Felizola Saulo J A; Ise Kazue; McNamara Keely M; Sasano Hironobu;
Ozawa Yohei; Inoue Takayoshi; Doi Masao; Okamura Hitoshi; et al
From The Journal of steroid biochemistry and molecular biology (2014), 144 Pt B268-79, Language: English,
Database: MEDLINE
The sebaceous gland is a major site of steroid synthesis in human skin, but details of the status of steroidogenic
enzymes and their regulation in human sebaceous glands under normal and pathological conditions have rarely been
reported. Therefore, in this study, we examined the status of steroidogenic enzymes, sex steroid receptors and
transcription factors in human sebaceous glands under normal and pathological conditions to explore their possible
roles in in situ steroid production in human skin. Immunohistochemical analysis was performed in a total of 59 human
skin specimens, including 22 normal human sebaceous glands, 12 with sebaceous nevus, 12 with sebaceous gland
hyperplasia, 3 with sebaceoma and 10 with sebaceous carcinoma. Immortalised human SZ95 sebocytes were treated
with forskolin or vehicle for 3h, 6h, 12h or 24h, and the mRNA levels of steroidogenic enzymes were evaluated at each
time point using quantitative RT-PCR (qPCR). The results of immunohistochemistry demonstrated the
immunoreactivity of 3β-HSD1, CYP11A1, StAR, 17β-HSD5, CYP17A1, 5α-red1, PRB, AR and NGFI-B in normal
human sebaceous gland, with lower levels of expression in pathological sebaceous glands. The results of the in vitro
study also indicated that the expression levels of 3β-HSD1, CYP11A1, StAR, 5α-red1 and NGFI-B were elevated by
forskolin. 3β-HSD1 and other steroidogenic enzymes were expressed in sebaceous glands resulting in in situ androgen
and progesterone synthesis and their functions.
~2 Citings
330. Ghrelin and des-acyl ghrelin inhibit aromatase expression and activity in human adipose stromal cells:
suppression of cAMP as a possible mechanism
By Docanto Maria M; Yang Fangyuan; Callaghan Brid; Au CheukMan C; Ragavan Rahini; Wang Xuyi; Furness John B;
Andrews Zane B; Brown Kristy A
Aromatase converts androgens into estrogens and its expression within adipose stromal cells (ASCs) is believed to be
the major driver of estrogen-dependent cancers in older women. Ghrelin is a gut-hormone that is involved in the
regulation of appetite and known to bind to and activate the cognate ghrelin receptor, GHSR1a. The unacylated form
of ghrelin, des-acyl ghrelin, binds weakly to GHSR1a but has been shown to play an important role in regulating a
number of physiological processes. The aim of this study was to determine the effect of ghrelin and des-acyl ghrelin
on aromatase in primary human ASCs. Primary human ASCs were isolated from adipose tissue of women undergoing
cosmetic surgery. Real-time PCR and tritiated water-release assays were performed to examine the effect of
treatment on aromatase transcript expression and aromatase activity, respectively. Treatments included ghrelin, des-
acyl ghrelin, obestatin, and capromorelin (GHSR1a agonist). GHSR1a protein expression was assessed by Western
blot and effects of treatment on Ca(2+) and cAMP second messenger systems were examined using the Flexstation
assay and the Lance Ultra cAMP kit, respectively. Results demonstrate that pM concentrations of ghrelin and des-acyl
ghrelin inhibit aromatase transcript expression and activity in ASCs under basal conditions and in PGE2-stimulated
cells. Moreover, the effects of ghrelin and des-acyl ghrelin are mediated via effects on aromatase promoter PII-specific
transcripts. Neither the GHSR1a-specific agonist capromorelin nor obestatin had any effect on aromatase transcript
expression or activity. Moreover, GHSR1a protein was undetectable by Western blot and neither ghrelin nor
capromorelin elicited a calcium response in ASCs. Finally, ghrelin caused a significant decrease in basal and
forskolin-stimulated cAMP in ASC. These findings suggest that ghrelin acts at alternate receptors in ASCs by
decreasing intracellular cAMP levels. Ghrelin mimetics may be useful in the treatment of estrogen-dependent breast
cancer.
~2 Citings
331. Enhancement of catecholamine release from PC12 cells by the traditional Japanese medicine, rikkunshito
By Nagamura Yuko; Terawaki Kiyoshi; Uezono Yasuhito; Tsukada Toshihiko
From BMC complementary and alternative medicine (2014), 14256, Language: English, Database: MEDLINE
BACKGROUND: Rikkunshito is a traditional Japanese herbal medicine that is used to treat appetite loss associated
with cancer and other disorders. The formulation contains various constituents that influence cell signaling, and
rikkunshito may accordingly affect human homeostasis through multiple regulatory pathways, including those governed
by the endocrine system. We investigated the actions of rikkunshito on catecholamine release from PC12 cells, an
adrenal chromaffin cell line. METHODS: The actions of rikkunshito on PC12 cells were evaluated by measuring
intracellular cAMP levels, tyrosine hydroxylase (TH) and vasoactive intestinal peptide (VIP) mRNA expression levels,
and catecholamine levels in the culture medium. The transcriptional activation of VIP gene by rikkunshito was
assessed by using a VIP promoter-driven reporter gene assay. RESULTS: Rikkunshito dose-dependently enhanced
forskolin-induced elevations in cAMP in PC12 cells, and also increased the gene expression of TH and VIP. The
transcriptional activation of VIP gene by rikkunshito was confirmed. Norepinephrine and dopamine secretion into the
culture medium of PC12 cells were also dose-dependently augmented by rikkunshito and/or forskolin, but experiments
with a protein kinase C (PKC) activator and a phosphodiesterase inhibitor revealed that the effects of rikkunshito were
not simply due to the modulation of PKC or phosphodiesterase activity. CONCLUSIONS: These findings suggest that
rikkunshito enhances the release of catecholamines by a novel mechanism involving cAMP.
~0 Citings
332. Hyperphosphorylation of PP2A in colorectal cancer and the potential therapeutic value showed by its
forskolin-induced dephosphorylation and activation
By Cristobal Ion; Garcia-Foncillas Jesus; Rincon Raul; Carames Cristina; del Puerto-Nevado Laura; Manso Rebeca;
Madoz-Gurpide Juan; Rojo Federico
From Biochimica et biophysica acta (2014), 1842(9), 1823-9, Language: English, Database: MEDLINE
BACKGROUND: The tumor suppressor protein phosphatase 2A (PP2A) is frequently inactivated in human cancer and
phosphorylation of its catalytic subunit (p-PP2A-C) at tyrosine-307 (Y307) has been described to inhibit this
phosphatase. However, its molecular and clinical relevance in colorectal cancer (CRC) remains unclear. METHODS:
p-PP2A-C Y307 was determined by immunoblotting in 7 CRC cell lines and 35 CRC patients. CRC cells were treated
with the PP2A activator forskolin alone or combined with the PP2A inhibitor okadaic acid, 5-fluorouracil and oxaliplatin.
We examined cell growth, colonosphere formation, caspase activity and AKT and ERK activation. RESULTS: PP2A-C
was found hyperphosphorylated in CRC cell lines. Forskolin dephosphorylated and activated PP2A, impairing
proliferation and colonosphere formation, and inducing activation of caspase 3/7 and changes in AKT and ERK
phosphorylation. Moreover, forskolin showed additive effects with 5-fluorouracil and oxaliplatin treatments. Analysis of
p-PP2A-C Y307 in primary tumors confirmed the presence of this alteration in a subgroup of CRC patients.
CONCLUSIONS: Our data show that PP2A-C hyperphosphorylation is a frequent event that contributes to PP2A
inhibition in CRC. Antitumoral effects of forskolin-mediated PP2A activation suggest that the analysis of p-PP2A-C
Y307 status could be used to identify a subgroup of patients who would benefit from treatments based on PP2A
activators.
~4 Citings
333. Growth arrest and forced differentiation of human primary glioblastoma multiforme by a novel small
molecule
By Kang Tae-Wook; Choi Soon Won; Yang Se-Ran; Shin Tae-Hoon; Kang Kyung-Sun; Kim Hyung-Sik; Yu Kyung-Rok;
Hong In-Sun; Ro Seonggu; Cho Joong Myung
From Scientific reports (2014), 45546, Language: English, Database: MEDLINE
Glioblastoma multiforme is the most common malignant brain tumor in adults, with an average survival of less than one
year due to its resistance to therapy. Recent studies reported that GBM initiates from CD133-expressing cancer stem
cells (CSC). However, the efficacy of CSC targeting is limited. A newly developed approach in cancer treatment is the
forced differentiation of cancer cells. Here, we show that the treatment of the novel small molecule, CG500354, into
CD133-expressing human primary GBM cells induces growth arrest by cell cycle regulators, p53, p21, p27 and phase-
specific cyclins, and neural differentiation, as confirmed by neural progenitor/precursor markers, nestin, GFAP and
Tuj1. When GBM-derived cells caused the tumors in NOD/SCID mice, CG500354 induced GBM-derived cells
differentiation into Tuj1 and GFAP expressing cells. We next demonstrated that CG500354 plays a tumor-suppressive
role via cAMP/CREB signaling pathway. CG500354 increases not only the extracellular cAMP level but also the
protein level of PKA and CREB. Additionally, both mimetic substances, Forskolin and Rolipram, revealed comparable
results with CG500354. Our findings indicate that induction of growth arrest and neural differentiation via cAMP/CREB
signaling pathway by CG500354 treatment suggests the novel targeting of PDE4D in the development of new drugs for
brain tumor therapy.
~6 Citings
334. Inhibitory effects of prostaglandin E2 on collagen synthesis and cell proliferation in human stellate cells
from pancreatic head adenocarcinoma
By Pomianowska Ewa; Sandnes Dagny; Grzyb Krzysztof; Schjolberg Aasa R; Aasrum Monica; Tveteraas Ingun H;
Tjomsland Vegard; Christoffersen Thoralf; Gladhaug Ivar P
From BMC cancer (2014), 14413, Language: English, Database: MEDLINE
BACKGROUND: Several studies have described an increased cyclooxygenase-2 (COX-2) expression in pancreatic
cancer, but the role of COX-2 in tumour development and progression is not clear. The aim of the present study was
to examine expression of COX-2 in cancer cells and stromal cells in pancreatic cancer specimens, and to explore the
role of PGE2 in pancreatic stellate cell proliferation and collagen synthesis. METHODS: Immunohistochemistry and
immunofluorescence was performed on slides from whole sections of tissue blocks using antibodies against COX-2
and α-smooth muscle actin (αSMA). Pancreatic stellate cells (PSC) were isolated from surgically resected tumour
tissue by the outgrowth method. Cells were used between passages 4 and 8. Collagen synthesis was determined by
[(3)H]-proline incorporation, or by enzyme immunoassay measurement of collagen C-peptide. DNA synthesis was
measured by incorporation of [(3)H]-thymidine in DNA. Cyclic AMP (cAMP) was determined by radioimmunoassay.
Collagen 1A1 mRNA was determined by RT-qPCR. RESULTS: Immunohistochemistry staining showed COX-2 in
pancreatic carcinoma cells, but not in stromal cells. All tumours showed positive staining for αSMA in the fibrotic
stroma. Cultured PSC expressed COX-2, which could be further induced by interleukin-1β (IL-1β), epidermal growth
factor (EGF), thrombin, and PGE2, but not by transforming growth factor-β1 (TGFβ). Indirect coculture with the
adenocarcinoma cell line BxPC-3, but not HPAFII or Panc-1, induced COX-2 expression in PSC. Treatment of PSC
with PGE2 strongly stimulated cAMP accumulation, mediated by EP2 receptors, and also stimulated phosphorylation
of extracellular signal-regulated kinase (ERK). Treatment of PSC with PGE2 or forskolin suppressed both TGFβ-
stimulated collagen synthesis and PDGF-stimulated DNA synthesis. CONCLUSIONS: The present results show that
COX-2 is mainly produced in carcinoma cells and suggest that the cancer cells are the main source of PGE2 in
pancreatic tumours. PGE2 exerts a suppressive effect on proliferation and fibrogenesis in pancreatic stellate cells.
These effects of PGE2 are mediated by the cAMP pathway and suggest a role of EP2 receptors.
~2 Citings
335. CFTR and Anoctamin 1 (ANO1) contribute to cAMP amplified exocytosis and insulin secretion in human and
murine pancreatic beta-cells
By Edlund Anna; Esguerra Jonathan L S; Wendt Anna; Flodstrom-Tullberg Malin; Eliasson Lena
From BMC medicine (2014), 1287, Language: English, Database: MEDLINE
BACKGROUND: Mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene lead to the
disease cystic fibrosis (CF). Although patients with CF often have disturbances in glucose metabolism including
impaired insulin release, no previous studies have tested the hypothesis that CFTR has a biological function in
pancreatic beta-cells. METHODS: Experiments were performed on islets and single beta-cells from human donors
and NMRI-mice. Detection of CFTR was investigated using PCR and confocal microscopy. Effects on insulin
secretion were measured with radioimmunoassay (RIA). The patch-clamp technique was used to measure ion channel
currents and calcium-dependent exocytosis (as changes in membrane capacitance) on single cells with high temporal
resolution. Analysis of ultrastructure was done on transmission electron microscopy (TEM) images. RESULTS: We
detected the presence of CFTR and measured a small CFTR conductance in both human and mouse beta-cells. The
augmentation of insulin secretion at 16.7 mM glucose by activation of CFTR by cAMP (forskolin (FSK) or GLP-1) was
significantly inhibited when CFTR antagonists (GlyH-101 and/or CFTRinh-172) were added. Likewise, capacitance
measurements demonstrated reduced cAMP-dependent exocytosis upon CFTR-inhibition, concomitant with a
decreased number of docked insulin granules. Finally, our studies demonstrate that CFTR act upstream of the
chloride channel Anoctamin 1 (ANO1; TMEM16A) in the regulation of cAMP- and glucose-stimulated insulin secretion.
CONCLUSION: Our work demonstrates a novel function for CFTR as a regulator of pancreatic beta-cell insulin
secretion and exocytosis, and put forward a role for CFTR as regulator of ANO1 and downstream priming of insulin
granules prior to fusion and release of insulin. The pronounced regulatory effect of CFTR on insulin secretion is
consistent with impaired insulin secretion in patients with CF.
~11 Citings
336. Ultraviolet radiation, aging and the skin: prevention of damage by topical cAMP manipulation
By Amaro-Ortiz Alexandra; Yan Betty; D'Orazio John A
From Molecules (Basel, Switzerland) (2014), 19(5), 6202-19, Language: English, Database: MEDLINE
Being the largest and most visible organ of the body and heavily influenced by environmental factors, skin is ideal to
study the long-term effects of aging. Throughout our lifetime, we accumulate damage generated by UV radiation. UV
causes inflammation, immune changes, physical changes, impaired wound healing and DNA damage that promotes
cellular senescence and carcinogenesis. Melanoma is the deadliest form of skin cancer and among the malignancies
of highest increasing incidence over the last several decades. Melanoma incidence is directly related to age, with
highest rates in individuals over the age of 55 years, making it a clear age-related disease. In this review, we will focus
on UV-induced carcinogenesis and photo aging along with natural protective mechanisms that reduce amount of
"realized" solar radiation dose and UV-induced injury. We will focus on the theoretical use of forskolin, a plant-derived
pharmacologically active compound to protect the skin against UV injury and prevent aging symptoms by up-regulating
melanin production. We will discuss its use as a topically-applied root-derived formulation of the Plectranthus barbatus
(Coleus forskolii) plant that grows naturally in Asia and that has long been used in various Aryuvedic teas and
therapeutic preparations.
~4 Citings
337. Protective effect of an antithyroid compound against γ-radiation-induced damage in human colon cancer
cells
By Perona Marina; Dagrosa Maria A; Pagotto Romina; Casal Mariana; Pignataro Omar; Pisarev Mario A; Juvenal
Guillermo J
From Radiation and environmental biophysics (2014), 53(3), 611-9, Language: English, Database: MEDLINE
We have previously reported the radioprotective effect of propylthiouracil (PTU) on thyroid cells. The aim of the
present study was to analyze whether tumor cells and normal cells demonstrate the same response to PTU. Human
colon carcinoma cells were irradiated with γ-irradiation with or without PTU. We evaluated the clonogenic survival,
intracellular reactive oxygen species levels, catalase, superoxide dismutase and glutathione peroxidase activities, and
apoptosis by nuclear cell morphology and caspase-3 activity assays. Cyclic AMP (cAMP) levels were measured by
radioimmunoassay. PTU treatment increased surviving cell fraction at 2 Gy (SF2) from 56.9 ± 3.6 in controls to 75.0 ±
3.5 (p < 0.05) and diminished radiation-induced apoptosis. In addition, we observed that the level of antioxidant
enzymes' activity was increased in cells treated with PTU. Moreover, pretreatment with PTU increased intracellular
levels of cAMP. Forskolin (p < 0.01) and dibutyryl cAMP (p < 0.05) mimicked the effect of PTU on SF2. Co-treatment
with H89, an inhibitor of protein kinase A, abolished the radioprotective effect of PTU. PTU reduces the toxicity of
ionizing radiation by increasing cAMP levels and also possibly through a reduction in apoptosis levels and in radiation-
induced oxidative stress damage. We therefore conclude that PTU protects both normal and cancer cells during
exposure to radiation in conditions mimicking the radiotherapy.
~0 Citings
338. Live cell imaging of in vitro human trophoblast syncytialization

By Wang Rui; Zheng Ru; Lu Xiaoyin; Dang Yan-Li; Li Yue; Wang Li-Juan; Zhu Cheng; Lin Hai-Yan; Wang Hongmei; Li
Weiwei
Human trophoblast syncytialization, a process of cell-cell fusion, is one of the most important yet least understood
events during placental development. Investigating the fusion process in a placenta in vivo is very challenging given
the complexity of this process. Application of primary cultured cytotrophoblast cells isolated from term placentas and
BeWo cells derived from human choriocarcinoma formulates a biphasic strategy to achieve the mechanism of
trophoblast cell fusion, as the former can spontaneously fuse to form the multinucleated syncytium and the latter is
capable of fusing under the treatment of forskolin (FSK). Live-cell imaging is a powerful tool that is widely used to
investigate many physiological or pathological processes in various animal models or humans; however, to our
knowledge, the mechanism of trophoblast cell fusion has not been reported using a live- cell imaging manner. In this
study, a live-cell imaging system was used to delineate the fusion process of primary term cytotrophoblast cells and
BeWo cells. By using live staining with Hoechst 33342 or cytoplasmic dyes or by stably transfecting enhanced green
fluorescent protein (EGFP) and DsRed2-Nuc reporter plasmids, we observed finger-like protrusions on the cell
membranes of fusion partners before fusion and the exchange of cytoplasmic contents during fusion. In summary, this
study provides the first video recording of the process of trophoblast syncytialization. Furthermore, the various live-cell
imaging systems used in this study will help to yield molecular insights into the syncytialization process during
placental development.
~2 Citings
339. Calcium-signaling components in rat insulinoma β-cells (INS-1) and pancreatic islets are differentially
influenced by melatonin
By Bazwinsky-Wutschke Ivonne; Muhlbauer Eckhard; Albrecht Elke; Peschke Elmar
From Journal of pineal research (2014), 56(4), 439-49, Language: English, Database: MEDLINE
The pineal secretory product melatonin exerts its influence on the insulin secretion of pancreatic islets by different
signaling pathways. The purpose of this study was to analyze the impact of melatonin on calcium-signaling
components under different conditions. In a transfected INS-1 cell line overexpressing the human MT2 receptor
(hMT2-INS-1), melatonin treatment induced even stronger depressive effects on calcium/calmodulin-dependent kinase
2d and IV (Camk2d, CamkIV) transcripts during 3-isobutyl-1-methylxanthine (IBMX) treatment than in normal INS-1
cells, indicating a crucial influence of melatonin receptor density on transcript-level regulation. In addition, melatonin
induced a significant downregulation of calmodulin (Calm1) in IBMX-treated hMT2-INS-1 cells. Long-term
administration of melatonin alone reduced CamkIV transcript levels in INS-1 cells; however, transcript levels of
Camk2d remained unchanged. The release of insulin was diminished under long-term melatonin treatment. The
impact of melatonin also involved reductions in CAMK2D protein during IBMX or forskolin treatments in INS-1 cells, as
measured by an enzyme-linked immunosorbent assay, indicating a functional significance of transcriptional changes in
pancreatic islets. Furthermore, analysis of melatonin receptor knockout mice showed that the transcript levels of
Camk2d, CamkIV, and Calm1 were differentially influenced according to the melatonin receptor subtype deleted. In
conclusion, this study provides evidence that melatonin has different impacts on the regulation of Calm1 and Camk.
These calcium-signaling components are known as participants in the calcium/calmodulin pathway, which plays an
important functional role in the modulation of the β-cell signaling pathways leading to insulin secretion.
~0 Citings
340. Different effects of adenylyl cyclase activators and phosphodiesterases inhibitors on cervical cancer (HeLa)
and breast cancer (MCF-7) cells proliferation
By Mahdian Davood; Shafiee-Nick Reza; Mousavi Seyed Hadi
From Toxicology mechanisms and methods (2014), 24(4), 307-14, Language: English, Database: MEDLINE
Breast and cervical cancers are the most common cancers in Iran and worldwide. Hormonal stimulation of cyclic
adenosine mono phosphate (cAMP) and the cAMP-dependent protein kinase PKA regulates cell growth by different
mechanism. cAMP can stimulate cell growth in many cell types while inhibiting cell growth in others. In some cell lines
have been shown that the proliferation of tumor cells is reduced by increasing cAMP in cells. In this study, we evaluate
growth arrest of selective PDE3 and non-selective PDE inhibitors, which lead to increase level of cAMP in cervical
(HeLa) and breast cancer (MCF7) cell lines have been studied. Cells were incubated with different concentrations of
selective, non-selective PDE inhibitors, beta adrenergic receptor agonist and direct stimulator of adenylyl cyclase. Cell
viability was quantitated by MTT assay. Apoptotic cells were determined using PI staining of DNA fragmentation by
flow cytometry (sub-G1 peak). Result showed that selective PDE inhibitors decreased cell viability in HeLa and MCF-7
cells as a time-dependent manner. Non-selective inhibitor and beta-adrenergic receptor agonist also decrease cell
viability but they are less powerful than selective PDE3 inhibitors. Forskolin had no effect in viability of cells. Analysis
of DNA fragmentation by flow cytometry showed apoptosis involved in selective PDE3 inhibitors induced toxicity in
HeLa cell. Thus, the growth inhibitory effects of selective PDE3 inhibitors are more effective than non-selective
inhibitor. Further studies are needed to investigate the mechanism of action is on the field.
~2 Citings
341. cAMP signaling inhibits radiation-induced ATM phosphorylation leading to the augmentation of apoptosis
in human lung cancer cells
By Cho Eun-Ah; Kim Eui-Jun; Kwak Sahng-June; Juhnn Yong-Sung
From Molecular cancer (2014), 1336, Language: English, Database: MEDLINE
BACKGROUND: The ataxia-telangiectasia mutated (ATM) protein kinase plays a central role in coordinating the
cellular response to radiation-induced DNA damage. cAMP signaling regulates various cellular responses including
metabolism and gene expression. This study aimed to investigate the mechanism through which cAMP signaling
regulates ATM activation and cellular responses to ionizing radiation in lung cancer cells. METHODS: Lung cancer
cells were transfected with constitutively active stimulatory G protein (GαsQL), and irradiated with γ-rays. The
phosphorylation of ATM and protein phosphatase 2A was analyzed by western blotting, and apoptosis was assessed
by western blotting, flow cytometry, and TUNNEL staining. The promoter activity of NF-κB was determined by dual
luciferase reporter assay. BALB/c mice were treated with forskolin to assess the effect in the lung tissue. RESULTS:
Transient expression of GαsQL significantly inhibited radiation-induced ATM phosphorylation in H1299 human lung
cancer cells. Treatment with okadaic acid or knock down of PP2A B56δ subunit abolished the inhibitory effect of Gαs
on radiation-induced ATM phosphorylation. Expression of GαsQL increased phosphorylation of the B56δ and PP2A
activity, and inhibition of PKA blocked Gαs-induced PP2A activation. GαsQL enhanced radiation-induced cleavage of
caspase-3 and PARP and increased the number of early apoptotic cells. The radiation-induced apoptosis was
increased by inhibition of NF-κB using PDTC or inhibition of ATM using KU55933 or siRNA against ATM. Pretreatment
of BALB/c mice with forskolin stimulated phosphorylation of PP2A B56δ, inhibited the activation of ATM and NF-κB,
and augmented radiation-induced apoptosis in the lung tissue. GαsQL expression decreased the nuclear levels of the
p50 and p65 subunits and NF-κB-dependent activity after γ-ray irradiation in H1299 cells. Pretreatment with
prostaglandin E2 or isoproterenol increased B56δ phosphorylation, decreased radiation-induced ATM phosphorylation
and increased apoptosis. CONCLUSIONS: cAMP signaling inhibits radiation-induced ATM activation by PKA-
dependent activation of PP2A, and this signaling mechanism augments radiation-induced apoptosis by reducing ATM-
dependent activation of NF-κB in lung cancer cells.
~8 Citings
342. Hydrogen sulfide inhibits A2A adenosine receptor agonist induced β-amyloid production in SH-SY5Y
neuroblastoma cells via a cAMP dependent pathway
By Nagpure Bhushan Vijay; Bian Jin-Song
Alzheimer's disease (AD) is the leading cause of senile dementia in today's society. Its debilitating symptoms are
manifested by disturbances in many important brain functions, which are influenced by adenosine. Hence,
adenosinergic system is considered as a potential therapeutic target in AD treatment. In the present study, we found
that sodium hydrosulfide (NaHS, an H2S donor, 100 µM) attenuated HENECA (a selective A2A receptor agonist, 10-
200 nM) induced β-amyloid (1-42) (Aβ42) production in SH-SY5Y cells. NaHS also interfered with HENECA-stimulated
production and post-translational modification of amyloid precursor protein (APP) by inhibiting its maturation.
Measurement of the C-terminal APP fragments generated from its enzymatic cleavage by β-site amyloid precursor
protein cleaving enzyme 1 (BACE1) showed that NaHS did not have any significant effect on β-secretase activity.
However, the direct measurements of HENECA-elevated γ-secretase activity and mRNA expressions of presenilins
suggested that the suppression of Aβ42 production in NaHS pretreated cells was mediated by inhibiting γ-secretase.
NaHS induced reductions were accompanied by similar decreases in intracellular cAMP levels and phosphorylation of
cAMP responsive element binding protein (CREB). NaHS significantly reduced the elevated cAMP and Aβ42
production caused by forskolin (an adenylyl cyclase, AC agonist) alone or forskolin in combination with IBMX (a
phosphodiesterase inhibitor), but had no effect on those caused by IBMX alone. Moreover, pretreatment with NaHS
significantly attenuated HENECA-elevated AC activity and mRNA expressions of various AC isoforms. These data
suggest that NaHS may preferentially suppress AC activity when it was stimulated. In conclusion, H2S attenuated
HENECA induced Aβ42 production in SH-SY5Y neuroblastoma cells through inhibiting γ-secretase via a cAMP
dependent pathway.
~3 Citings
343. cAMP-PKA inhibition of SK3 channel reduced both Ca2+ entry and cancer cell migration by regulation of
SK3-Orai1 complex
By Clarysse Lucie; Gueguinou Maxime; Potier-Cartereau Marie; Vandecasteele Gregoire; Bougnoux Philippe;
Chevalier Stephan; Chantome Aurelie; Vandier Christophe
From Pflugers Archiv : European journal of physiology (2014), 466(10), 1921-32, Language: English, Database:
MEDLINE
SK3 channel mediates the migration of various cancer cells. When expressed in breast cancer cells, SK3 channel
forms a complex with Orai1, a voltage-independent Ca(2+) channel. This SK3-Orai1 complex associates within lipid
rafts where it controls a constitutive Ca(2+) entry leading to cancer cell migration and bone metastases development.
Since cAMP was found to modulate breast cancer cell migration, we hypothesized that this could be explained by a
modulation of SK3 channel activity. Herein, we study the regulation of SK3 channel by the cAMP-PKA pathway and
the consequences for SK3-dependent Ca(2+) entry and cancer cell migration. We established that the beta-adrenergic
receptor agonist, isoprenaline, or the direct adenylyl cyclase activator forskolin alone or in combination with the PDE4
inhibitor, CI-1044, decreased SK3 channel activity without modifying the expression of SK3 protein at the plasma
membrane. Forskolin and CI-1044 reduced the SK3-dependent constitutive Ca(2+) entry and the SK3-dependent
migration of MDA-MB-435s cells. PKA inhibition with KT 5720 reduced: (1) the effect of forskolin and CI-1044 by 50 %
on Ca(2+) entry and (2) SK3 activity by inhibiting the serine phosphorylation of SK3. These cAMP-elevating agents
displaced Orai1 protein outside lipid rafts in contrast to SK3, which remained in the lipid rafts fractions. All together,
these results show that activation of the cAMP-PKA pathway decreases SK3 channel and SK3-Orai1 complex
activities, leading to a decrease in both Ca(2+) entry and cancer cell migration. This work supports the potential use of
cAMP-elevating agents to reduce cancer cell migration and may provide novel opportunities to address/prevent bone
metastasis.
~3 Citings
344. Expression of mRNAs of Urocortin in the STKM-1 gastric cancer cell line
By Akiyoshi Kohei; Kamada Minori; Fujioka Kouki; Ikeda Keiichi; Tojo Katsuyoshi; Manome Yoshinobu
From Anticancer research (2013), 33(12), 5289-94, Language: English, Database: MEDLINE
BACKGROUND: Urocortin is analogous to corticotrophin-releasing factors (CRFs) and a member of the CRF family.
We previously demonstrated that urocortin mRNAs were expressed in both human and rat glioma cell lines, and that
some of these lines transcribed the receptors. We hypothesize that urocortin might also be expressed in a gastric
cancer cell line. The aim of the present study was to clarify the expression of mRNAs of urocortin1 (UCN1), -2 and -3
and of CRF and CRF receptors 1 and 2 in a gastric cancer cell line. MATERIALS AND METHODS: STKM-1 a poorly-
differentiated adenocarcinoma cell line was used. Transcripts in the cells were analyzed using cDNA. The fluctuation
of mRNA with cellular stress, such as the one caused by a chemotherapeutic agent, serum supplementation and
forskolin was examined. RESULTS: Transcripts of UCN1, -2 and CRFR2 were expressed. No changes in
transcription of UCN1 and UCN2 were observed with cellular stress. However, expression of CRFR2 mRNA
transcripts significantly increased after an initial 24-h exposure to forskolin. CONCLUSION: Expression of the mRNAs
of UCN1, 2 and CRFR2 was confirmed in the human gastric cancer cell line, STKM-1. Although the quantity of CRFR2
transcripts varied with forskolin, the overall transcription pattern was not influenced by cellular stimuli.
~0 Citings
345. Metallothionein-3 (MT-3) in the human adrenal cortex and its disorders
By Felizola Saulo J A; Nakamura Yasuhiro; Arata Yuki; Ise Kazue; Satoh Fumitoshi; Rainey William E; Midorikawa
Sanae; Suzuki Shinichi; Sasano Hironobu
From Endocrine pathology (2014), 25(3), 229-35, Language: English, Database: MEDLINE
Metallothionein-3 (MT-3) is an intracellular, low molecular weight protein mainly distributed in the central nervous
system but also in various peripheral organs and several types of human neoplasms. However, details of MT-3
expression have not been examined in human adrenal cortex and its disorders. The mRNA levels of MT-3 were first
evaluated by quantitative RT-PCR (qPCR) in adrenocortical aldosterone-producing adenoma (APA: 11) and cortisol-
producing adenoma (CPA: 14). In addition, MT-3 immunohistochemistry was performed in non-pathological adrenal
glands (NA: 19), idiopathic hyperaldosteronism (IHA: 10), APA (20), CPA (24), adjacent non-neoplastic adrenal glands
of adenoma (AAG: 20), and adrenocortical carcinoma (ACC: 8). H295R cells were also treated with angiotensin-II or
forskolin in a time-dependent manner, and the changes of MT-3 mRNA levels were evaluated by qPCR. Results of
qPCR analysis demonstrated that MT-3 mRNA levels were significantly higher in APA than CPA (P = 0.0004). MT-3
immunoreactivity was detected in the zona glomerulosa of NA, IHA, and AAG, as well as in APA, CPA, and ACC.
When treated with angiotensin-II and forskolin, MT-3 mRNA levels reached a peak by 12 h in H295R cells, with
significantly higher levels compared to control non-treated cells (P < 0.01). The presence of MT-3 in the ZG of NA,
IHA, and AAG, as well as APA may imply a role in the pathophysiology of aldosterone-producing tissues.
~1 Citing

346. Cyclic AMP enhances progesterone action in human myometrial cells
By Chen Li; Lei Kaiyu; Malawana Johann; Yulia Angela; Sooranna Suren R; Bennett Phillip R; Liang Zhiqing;
Grammatopoulos Dimitri; Johnson Mark R
From Molecular and cellular endocrinology (2014), 382(1), 334-43, Language: English, Database: MEDLINE
Cyclic AMP (cAMP) has been shown to promote progesterone and glucocorticoid action in a variety of cellular settings.
In this study, we have used human myometrial cells to investigate whether cAMP potentiates the ability of
progesterone to repress IL-1β-driven COX-2 expression. We found that forskolin enhanced progesterone-repression
of IL-1β-driven COX-2 expression in association with delayed IL-1β-induced nuclear phospho-p65 entry and reduced
NF-κB binding to the COX-2 promoter. Further, forskolin enhanced the progesterone-induced expression of FKBP5
and 11βHSD1, progesterone-driven activity of a progesterone response element (PRE) and progesterone receptor
(PR)-B binding to a transfected PRE. In addition, forskolin treatment increased PR-B levels and reduced the PR-A:PR-
B ratio while acutely decreasing the association between PR and nuclear receptor co-repressor (NCoR) and reducing
NCoR levels after 6h. These findings are of importance in situations where enhancing progesterone activity is
desirable, for example in the management of endometrial cancer, the promotion of endometrial receptivity or the
maintenance of myometrial quiescence during pregnancy.
~3 Citings
347. High-content pSTAT3/1 imaging assays to screen for selective inhibitors of STAT3 pathway activation in
head and neck cancer cell lines
By Johnston Paul A; Sen Malabika; Hua Yun; Camarco Daniel; Shun Tong Ying; Lazo John S; Grandis Jennifer R
From Assay and drug development technologies (2014), 12(1), 55-79, Language: English, Database: MEDLINE
The oncogenic transcription factor signal transducer and activator of transcription 3 (STAT3) is hyperactivated in most
cancers and represents a plausible therapeutic target. In the absence of STAT3-selective small-molecule inhibitors,
we sought to develop pSTAT3/1 high-content imaging (HCS) assays to screen for selective inhibitors of STAT3
pathway activation in head and neck squamous cell carcinomas (HNSCC) tumor cell lines. Based on the expression of
the interleukin-6 (IL-6)Rα and gp130 subunits of the IL-6 receptor complex and STAT3, we selected the Cal33 HNSCC
cell line as our model. After developing image acquisition and analysis procedures, we rigorously investigated the
cytokine activation responses to optimize the dynamic ranges of both assays and demonstrated that the pan-Janus
kinase inhibitor pyridone 6 nonselectively inhibited pSTAT3 and pSTAT1 activation with 50% inhibition concentrations
of 7.19 ± 4.08 and 16.38 ± 8.45 nM, respectively. The optimized pSTAT3 HCS assay performed very well in a pilot
screen of 1,726 compounds from the Library of Pharmacologically Active Compounds and the National Institutes of
Health clinical collection sets, and we identified 51 inhibitors of IL-6-induced pSTAT3 activation. However, only three
of the primary HCS actives selectively inhibited STAT3 compared with STAT1. Our follow-up studies indicated that the
nonselective inhibition of cytokine induced pSTAT3 and pSTAT1 activation by G-alpha stimulatory subunit-coupled G-
protein-coupled receptor agonists, and forskolin was likely due to cyclic adenosine monophosphate-mediated up-
regulation of suppressors of cytokine signaling 3. Azelastine, an H1 receptor antagonist approved for the treatment of
seasonal allergic rhinitis, nonallergic vasomotor rhinitis, and ocular conjunctivitis, was subsequently confirmed as a
selective inhibitor of IL-6-induced pSTAT3 activation that also reduced the growth of HNSCC cell lines. These data
illustrate the power of a chemical biology approach to lead generation that utilizes fully developed and optimized HCS
assays as phenotypic screens to interrogate specific signaling pathways.
~10 Citings

By Iancu Cristina V; Zamoon Jamillah; Woo Sang Bum; Aleshin Alexander; Choe Jun-yong
From Proceedings of the National Academy of Sciences of the United States of America (2013), 110(44), 17862-7,
Language: English, Database: MEDLINE
protein and lipid synthesis. These transporters are involved in important common diseases such as cancer and
diabetes. Here, we report the crystal structure of the Staphylococcus epidermidis glucose/H(+) symporter in an inward-
facing conformation at 3.2-ÅA resolution. The Staphylococcus epidermidis glucose/H(+) symporter is homologous to
human glucose transporters, is very specific and has high avidity for glucose, and is inhibited by the human glucose
transport inhibitors cytochalasin B, phloretin, and forskolin. On the basis of the crystal structure in conjunction with
mutagenesis and functional studies, we propose a mechanism for glucose/H(+) symport and discuss the symport
mechanism versus facilitated diffusion.
~22 Citings
349. Cyclic AMP regulates the migration and invasion potential of human pancreatic cancer cells
By Zimmerman Noah P; Roy Ishan; Hauser Andrew D; Wilson Jessica M; Williams Carol L; Dwinell Michael B
From Molecular carcinogenesis (2015), 54(3), 203-15, Language: English, Database: MEDLINE
Aggressive dissemination and metastasis of pancreatic ductal adenocarcinoma (PDAC) results in poor prognosis and
marked lethality. Rho monomeric G protein levels are increased in pancreatic cancer tissue. As the mechanisms
underlying PDAC malignancy are little understood, we investigated the role for cAMP in regulating monomeric G
protein regulated invasion and migration of pancreatic cancer cells. Treatment of PDAC cells with cAMP elevating
agents that activate adenylyl cyclases, forskolin, protein kinase A (PKA), 6-Bnz-cAMP, or the cyclic nucleotide
phosphodiesterase inhibitor cilostamide significantly decreased migration and Matrigel invasion of PDAC cell lines.
Inhibition was dose-dependent and not significantly different between forskolin or cilostamide treatment. cAMP
elevating drugs not only blocked basal migration, but similarly abrogated transforming-growth factor-β-directed PDAC
cell migration and invasion. The inhibitory effects of cAMP were prevented by the pharmacological blockade of PKA.
Drugs that increase cellular cAMP levels decreased levels of active RhoA or RhoC, with a concomitant increase in
phosphorylated RhoA. Diminished Rho signaling was correlated with the appearance of thickened cortical actin bands
along the perimeter of non-motile forskolin or cilostamide-treated cells. Decreased migration did not reflect alterations
in cell growth or programmed cell death. Collectively these data support the notion that increased levels of cAMP
specifically hinder PDAC cell motility through F-actin remodeling.
~10 Citings
350. Hedgehog-signaling is upregulated in non-producing human adrenal adenomas and antagonism of

hedgehog-signaling inhibits proliferation of NCI-H295R cells and an immortalized primary human adrenal cell
line
By Werminghaus Pascal; Haase Matthias; Hornsby Peter J; Schinner Sven; Schott Matthias; Malendowicz Ludwik K;
Lammers Bernhard J; Goretzki Peter E; Muller-Mattheis Volker; Markus Giessing; et al
MEDLINE
Hedgehog (Hh)-signaling pathway is important in embryonic development. Activation of Hh-signaling is associated
with tumorigenesis. Recent studies demonstrate that Hh-signaling is involved in the development of the adrenal gland
in mice and is important in regulating adrenal proliferation. We studied the expression of Sonic hedgehog (SHH),
Smoothened (SMO), Patched1 (PTCH1) and GLI family zinc finger 1 (GLI1) in human adrenal and in adrenocortical
tumors using immunohistochemistry and semi-quantitative reverse transcriptase-polymerase chain reaction.
Modulation of GLI1 and SMO messenger ribonucleic acid (mRNA) expression was investigated with forskolin. The role
of Hh-signaling was studied in NCI-H295R cells and in an immortalized primary cell line using the Hh-agonist
smoothened agonist (SAG) and the Hh-antagonist cyclopamine. The Hh-pathway components SHH, GLI1, PTCH1
and SMO were detectable in all adrenal glands. While in cortisol-producing adenomas (CPA), Hh-signaling expression
levels were comparable to that in normal adrenal cortex, a much higher mRNA expression of GLI1, SMO and SHH was
observed in non-producing adenomas (NPA). Interestingly, stimulation of cultured adrenal cells with forskolin led to a
decrease in expression of GLI1 and SMO mRNAs. Antagonism of Hh-signaling resulted in a lower proliferation rate of
adrenocortical cells, while Hh-agonism had no significant effect on adrenal cell proliferation. Our data show Hh-
signaling activity in adult adrenal glands. Activation of the PKA pathway results in lower expression of Hh-signaling
proteins. This might explain the lower expression of the Hh components GLI1 and SMO in CPA in comparison to the
higher expression in NPA. Hh-signaling might be involved in the tumorigenesis of NPA.
~2 Citings
351. Pharmacologic induction of epidermal melanin and protection against sunburn in a humanized mouse
model
By Amaro-Ortiz Alexandra; Vanover Jillian C; Scott Timothy L; D'Orazio John A
From Journal of visualized experiments : JoVE (2013), (79), , Language: English, Database: MEDLINE
Fairness of skin, UV sensitivity and skin cancer risk all correlate with the physiologic function of the melanocortin 1
receptor, a Gs-coupled signaling protein found on the surface of melanocytes. Mc1r stimulates adenylyl cyclase and
cAMP production which, in turn, up-regulates melanocytic production of melanin in the skin. In order to study the
mechanisms by which Mc1r signaling protects the skin against UV injury, this study relies on a mouse model with
"humanized skin" based on epidermal expression of stem cell factor (Scf). K14-Scf transgenic mice retain
melanocytes in the epidermis and therefore have the ability to deposit melanin in the epidermis. In this animal model,
wild type Mc1r status results in robust deposition of black eumelanin pigment and a UV-protected phenotype. In
contrast, K14-Scf animals with defective Mc1r signaling ability exhibit a red/blonde pigmentation, very little eumelanin
in the skin and a UV-sensitive phenotype. Reasoning that eumelanin deposition might be enhanced by topical agents
that mimic Mc1r signaling, we found that direct application of forskolin extract to the skin of Mc1r-defective fair-skinned
mice resulted in robust eumelanin induction and UV protection (1). Here we describe the method for preparing and
applying a forskolin-containing natural root extract to K14-Scf fair-skinned mice and report a method for measuring UV
sensitivity by determining minimal erythematous dose (MED). Using this animal model, it is possible to study how
epidermal cAMP induction and melanization of the skin affect physiologic responses to UV exposure.
~3 Citings
352. Red blood cell aggregation changes are depanded on its initial value: Effect of long-term drug treatment
By Muravyov A V; Tikhomirova I A; Maimistova A A; Bulaeva S V; Mikhailov P V; Kislov N V
From Clinical hemorheology and microcirculation (2012), , Language: English, Database: MEDLINE
This study was designed to investigate whether the red cell aggregation depends on its initial level under drug therapy
or cell incubation with bioactive chemical compounds. Sixty six subjects were enrolled onto this study, and sub-divided
into two groups: the first group of patients (n = 36) with cerebral atherosclerosis received pentoxifylline therapy (400
mg, thrice daily) for 4 weeks. The patients of the second group were initially treated with Epoetin beta 10,000 units
subcutaneously thrice a week, for 4 weeks. The second group - adult anemic patients (n = 30) with the confirmed
diagnosis of solid cancer (Hb < 100 g/L). After 4 weeks of pentoxifylline treatment the red cell aggregation increased
(p < 0.05) in the patients with initially low RBCA. On the other hand in the patients with initially high RBCA treatment
with pentoxifylline reduced it markedly (p < 0.01). In vitro experiments with pentoxifylline RBC incubation resulted in a
decrease of the initially high RBCA by 47% (p < 0.01), whereas in the sub-group with initially low RBCA it increased. It
was observed that after 4 weeks of epoetin-beta treatment 75% the anemic patients with initially high RBCA had an
aggregation lowering. The drop of aggregation was about 34% (p < 0.01). At the same time 25% of the study patients
had a significant RBCA increase (p < 0.05) after treatment. The initially low red cell aggregation after incubation with
epoetin-beta was markedly increased by 122% (p < 0.05). On the contrary initially high RBCA was reduced by 47% (p
< 0.05). When forskolin (10 µM) was added to the RBC suspensions the RBCA was increased in sub-group of
subjects with initially low aggregation and it was decreased in sub-group with initially high one. The similar RBCA
changes were observed when RBC suspensions were incubated with vinpocetine, calcium ionophore (A23187),
Phorbol 12-myristate 13-acetate (PMA) as a protein kinase C (PKC) stimulator. A major finding of this study is that the
red cell aggregation effects of some drugs depend markedly on the initial, pre-treatment aggregation status of the
patients. These results demonstrate that the different red blood cell aggregation responses to the biological stimuli
depend strongly on the initial, pre-treatment status of the subject and the most probably it is connected with the
crosstalk between the adenylyl cyclase signaling pathway and Ca2+ regulatory mechanism.
~0 Citings
353. Hedgehog signal inhibitors suppress the invasion of human rhabdomyosarcoma cells
By Oue Takaharu; Uehara Shuichiro; Yamanaka Hiroaki; Nomura Motonari; Usui Noriaki
From Pediatric surgery international (2013), 29(11), 1153-8, Language: English, Database: MEDLINE
PURPOSE: In the treatment of rhabdomyosarcoma (RMS), invasion and metastasis remain the most critical
determinants of resectability and survival. The objective of this study was to determine whether Hedgehog (Hh)
signaling plays a role in the invasion of RMS. METHODS: Two kinds of specific Hh signaling inhibitors, cyclopamine
and forskolin, were used to suppress activated Hh signals in three RMS cell lines. The effects of the Hh signaling
inhibitors on tumor cell invasion and motility were investigated using Matrigel invasion assays and wound closure
assays, respectively. RESULTS: The number of invaded cells counted in six random microscopic fields in the Matrigel
chambers was significantly decreased by both cyclopamine and forskolin in every RMS cell line. Furthermore, the
wound closure assays revealed that a blockade of the Hh signaling pathway by the Hh inhibitors strongly impairs RMS
cell motility, as visualized by the delayed closure of the gaps generated in the cultured cell monolayers of the three
RMS cell lines. CONCLUSIONS: Both the invasive capacity and motility of RMS cells are significantly suppressed by
Hh signaling inhibitors, demonstrating that the Hh pathway plays an important role in the invasion of RMS. Hh
inhibitors may provide a new paradigm for the treatment of RMS.
~0 Citings
354. Involvement of spinal PKA/CREB signaling pathway in the development of bone cancer pain
By Hang Li-Hua; Yang Jian-Ping; Shao Dong-Hua; Chen Zheng; Wang Hong
From Pharmacological reports : PR (2013), 65(3), 710-6, Language: English, Database: MEDLINE
BACKGROUND: It has been shown that spinal PKA/CREB signaling pathway is involved in neuropathic and
inflammatory pain, but its effects on bone cancer pain have not previously been investigated. The aim of this study
was to examine the potential role of the spinal PKA/CREB signaling pathway in the development of bone cancer pain.
METHODS: A bone cancer pain model was made by inoculation of Walker 256 cells into the intramedullary space of
rat tibia. Western blot analysis examined the expression of PKAca (PKA catalytic subunit) and phospho-CREB (p-
CREB) protein levels. The authors further investigated effects of intrathecal treatment with H-89 (a PKA inhibitor, 8
nmol) or forskolin (a PKA agonist, 10 nmol) on nociceptive behavior and the expression of PKAca and p-CREB.
RESULTS: On days 6, 9, and 15 after inoculation, the expression of PKAca and p-CREB protein levels were higher in
the bone cancer pain rats compared to the sham rats. On day 9, intrathecal administration of H-89 significantly
attenuated bone cancer-induced mechanical allodynia as well as upregulation of PKAca and p-CREB protein levels.
These effects were completely abolished by intrathecal pretreatment with the PKA agonist forskolin. CONCLUSION:
The results suggest that the spinal PKA/CREB signaling pathway may participate in the development of bone cancer
pain. The findings of this study may provide an evidence for developing novel analgesics to block bone cancer pain.
~1 Citing
355. Protein kinase C-induced activin A switches adrenocortical steroidogenesis to aldosterone by suppressing
CYP17A1 expression
By Hofland Johannes; Steenbergen Jacobie; Hofland Leo J; van Koetsveld Peter M; Eijken Marco; van Nederveen
Francien H; Kazemier Geert; de Herder Wouter W; Feelders Richard A; de Jong Frank H
From American journal of physiology. Endocrinology and metabolism (2013), 305(6), E736-44, Language: English,
Database: MEDLINE
Functional zonation of the adrenal cortex is a consequence of the zone-specific expression of P450c17 (CYP17A1)
and its cofactors. Activin and inhibin peptides are differentially produced within the zones of the adrenal cortex and
have been implicated in steroidogenic control. In this study, we investigated whether activin and inhibin can function
as intermediates in functional zonation of the human adrenal cortex. Activin A suppressed CYP17A1 expression and
P450c17 function in adrenocortical cell lines as well as in primary adrenal cell cultures. Inhibin βA-subunit mRNA and
activin A protein levels were found to be increased up to 1,900-fold and 49-fold, respectively, after protein kinase C
(PKC) stimulation through PMA or angiotensin II in H295R adrenocortical carcinoma cells. This was confirmed in
HAC15 cells and for PMA in primary adrenal cell cultures. Both PMA and Ang II decreased CYP17A1 expression in
the adrenocortical cell lines, whereas PMA concurrently suppressed CYP17A1 levels in the primary cultures. Inhibition
of activin signaling during PKC stimulation through silencing of the inhibin βA-subunit or blocking of the activin type I
receptor opposed the PMA-induced downregulation of CYP17A1 expression and P450c17 function. In contrast, PKA
stimulation through adrenocorticotrophin or forskolin increased expression of the inhibin α-subunit and betaglycan,
both of which are antagonists of activin action. These data indicate that activin A acts as a PKC-induced paracrine
factor involved in the suppression of CYP17A1 in the zona glomerulosa and can thereby contribute to functional
adrenocortical zonation.
~3 Citings
356. Interaction of PACAP with Sonic hedgehog reveals complex regulation of the hedgehog pathway by PKA
By Niewiadomski Pawel; Zhujiang Annie; Youssef Mary; Waschek James A
From Cellular signalling (2013), 25(11), 2222-30, Language: English, Database: MEDLINE
Sonic hedgehog (Shh) signaling is essential for proliferation of cerebellar granule cell progenitors (cGCPs) and its
aberrant activation causes a cerebellar cancer medulloblastoma. Pituitary adenylate cyclase activating polypeptide
(PACAP) inhibits Shh-driven proliferation of cGCPs and acts as tumor suppressor in murine medulloblastoma. We
show that PACAP blocks canonical Shh signaling by a mechanism that involves activation of protein kinase A (PKA)
and inhibition of the translocation of the Shh-dependent transcription factor Gli2 into the primary cilium. PKA is shown
to play an essential role in inhibiting gene transcription in the absence of Shh, but global PKA activity levels are found
to be a poor predictor of the degree of Shh pathway activation. We propose that the core Shh pathway regulates a
small compartmentalized pool of PKA in the vicinity of primary cilia. GPCRs that affect global PKA activity levels, such
as the PACAP receptor, cooperate with the canonical Shh signal to regulate Gli protein phosphorylation by PKA. This
interaction serves to fine-tune the transcriptional and physiological function of the Shh pathway.
~16 Citings
357. Bronchorelaxation of the human bronchi by CFTR activators

By Norez Caroline; Becq Frederic; Jayle Christophe; Vandebrouck Clarisse
From Pulmonary pharmacology & therapeutics (2014), 27(1), 38-43, Language: English, Database: MEDLINE
The airway functions are profoundly affected in many diseases including asthma, COPD and cystic fibrosis (CF). CF
the most common lethal autosomal recessive genetic disease is caused by mutations of the CFTR (Cystic Fibrosis
transmembrane Conductance Regulator) gene, which normally encodes a multifunctional and integral membrane
cAMP regulated and ATP gated Cl(-) channel expressed in airway epithelial cells. Using human lung tissues obtained
from patients undergoing surgery for lung cancer, we demonstrated that CFTR participates in bronchorelaxation.
Using human bronchial smooth muscle cells (HBSMC), we applied iodide influx assay to analyze the CFTR-dependent
ionic transport and immunofluorescence technique to localize CFTR proteins. Moreover, the relaxation was studied in
isolated human bronchial segments after pre-contraction with carbachol to determine the implication of CFTR in
bronchodilation. We found in HBSMC that the pharmacology and regulation of CFTR is similar to that of its epithelial
counterpart both for activation (using forskolin/genistein or a benzo[c]quinolizinium derivative) and for inhibition
(CFTR(inh)-172 and GPinh5a). With human bronchial rings, we observed that whatever the compound used including
salbutamol, the activation of muscular CFTR leads to a bronchodilation after constriction with carbachol. Altogether,
these observations revealed that CFTR in the human airways is expressed in bronchial smooth muscle cells and can
be pharmacologically manipulated leading to the hypothesis that this ionic channel could contribute to bronchodilation
in human.
~4 Citings
358. cAMP inhibits migration, ruffling and paxillin accumulation in focal adhesions of pancreatic ductal
adenocarcinoma cells: effects of PKA and EPAC
By Burdyga Alex; Conant Alan; Haynes Lee; Zhang Jin; Jalink Kees; Sutton Robert; Neoptolemos John; Costello
Eithne; Tepikin Alexei
From Biochimica et biophysica acta (2013), 1833(12), 2664-2672, Language: English, Database: MEDLINE
We demonstrated that increasing intracellular cAMP concentrations result in the inhibition of migration of PANC-1 and
other pancreatic ductal adenocarcinoma (PDAC) cell types. The rise of cAMP was accompanied by rapid and
reversible cessation of ruffling, by inhibition of focal adhesion turnover and by prominent loss of paxillin from focal
adhesions. All these phenomena develop rapidly suggesting that cAMP effectors have a direct influence on the
cellular migratory apparatus. The role of two primary cAMP effectors, exchange protein activated by cAMP (EPAC)
and protein kinase A (PKA), in cAMP-mediated inhibition of PDAC cell migration and migration-associated processes
was investigated. Experiments with selective activators of EPAC and PKA demonstrated that the inhibitory effect of
cAMP on migration, ruffling, focal adhesion dynamics and paxillin localisation is mediated by PKA, whilst EPAC
potentiates migration.
~15 Citings
359. Mitochondrial complex I inhibitors and forced oxidative phosphorylation synergize in inducing cancer cell
death
By Palorini Roberta; Simonetto Tiziana; Cirulli Claudia; Chiaradonna Ferdinando
From International journal of cell biology (2013), 2013243876, Language: English, Database: MEDLINE
Cancer cells generally rely mostly on glycolysis rather than oxidative phosphorylation (OXPHOS) for ATP production.
In fact, they are particularly sensitive to glycolysis inhibition and glucose depletion. On the other hand mitochondrial
dysfunctions, involved in the onset of the Warburg effect, are sometimes also associated with the resistance to
apoptosis that characterizes cancer cells. Therefore, combined treatments targeting both glycolysis and mitochondria
function, exploiting peculiar tumor features, might be lethal for cancer cells. In this study, we show that glucose
deprivation and mitochondrial Complex I inhibitors synergize in inducing cancer cell death. In particular, our results
reveal that low doses of Complex I inhibitors, ineffective on immortalized cells and in high glucose growth, become
specifically cytotoxic on cancer cells deprived of glucose. Importantly, the cytotoxic effect of the inhibitors on cancer
cells is strongly enhanced by forskolin, a PKA pathway activator, that we have previously shown to stimulate OXPHOS.
Taken together, we demonstrate that induction in cancer cells of a switch from a glycolytic to a more respirative
metabolism, obtained by glucose depletion or mitochondrial activity stimulation, strongly increases their sensitivity to
low doses of mitochondrial Complex I inhibitors. Our findings might be a valuable approach to eradicate cancer cells.
~6 Citings
360. 11β-hydroxyandrostenedione, the product of androstenedione metabolism in the adrenal, is metabolized in

LNCaP cells by 5α-reductase yielding 11β-hydroxy-5α-androstanedione
By Swart Amanda C; Schloms Lindie; Storbeck Karl-Heinz; Bloem Liezl M; Toit Therina du; Quanson Jonathan L;
Rainey William E; Swart Pieter
MEDLINE
11β-Hydroxyandrostenedione (11OHA4), which is unique to the adrenal, was first isolated from human adrenal tissue
in the fifties. It was later shown in the sixties that 11β-hydroxytestosterone (11OHT) was also produced by the human
adrenal. Attention has shifted back to these adrenal androgens once more, as improved analytical techniques have
enabled more accurate detection of steroid hormones. In this paper, we investigated the origin of these metabolites as
well as their subsequent metabolism and examined a possible physiological role for 11OHA4 in prostate cancer cells.
In H295R cells treated with forskolin and trilostane, etomidate, a reported cytochrome P450 11β-hydroxylase
(CYP11B1) inhibitor, blocked the production of corticosterone, cortisol, 11OHA4 and 11OHT. The metabolism of
androstenedione and testosterone by CYP11B1 and aldosterone synthase (CYP11B2) was assayed.
Androstenedione was converted by CYP11B1, while the conversion by CYP11B2 was negligible. Both enzymes
readily converted testosterone. The metabolism of these 11β-hydroxylated metabolites by 11β-hydroxysteroid
dehydrogenase (11βHSD) types 1 and 2 was subsequently investigated. 11βHSD2 catalyzed the conversion of both
11OHA4 and 11OHT to their respective keto-steroids, while 11βHSD1 catalyzed the conversion of 11-
ketoandrostenedione and 11-ketotestosterone to their respective hydroxy-steroids in Chinese hamster ovary cells.
Investigating a functional role, steroid 5α-reductase types 1 and 2 converted 11OHA4 to 11β-hydroxy-5α-
androstanedione (11OH-5α-dione), identified by accurate mass detection. UPLC-MS/MS analyses of 11OHA4
metabolism in LNCaP androgen-dependent prostate cancer cells, identified the 5α-reduced metabolite as well as 11-
ketoandrostenedione and 11-ketotestosterone, with the latter indicating conversion by 17β-hydroxysteroid
dehydrogenase. Downstream metabolism by 11βHSD2 and by 5α-reductase may therefore indicate a physiological
role for 11OHA4 and/or 11OH-5α-dione in normal and prostate cancer cells.
~3 Citings
361. Increased Th17 cells in the tumor microenvironment is mediated by IL-23 via tumor-secreted prostaglandin
E2
By Qian Xuesong; Gu Ling; Ning Huan; Zhang Yanping; Hsueh Eddy C; Fu Mingui; Hu Xiaoyu; Wei Lin; Hoft Daniel F;
Liu Jianguo
From Journal of immunology (Baltimore, Md. : 1950) (2013), 190(11), 5894-902, Language: English, Database:
MEDLINE
Tumor cell-derived molecules such as cytokines and lipid mediators play a critical role in inducing chronic inflammation
in the tumor microenvironment. We found that Th17 cells were increased in the peripheral blood, spleen, and tumor
tissues of mammary gland tumor-bearing mice. The Th17 cell survival factor, IL-23, was also overexpressed in tumor
tissues isolated from mice and human breast cancer patients. Soluble molecules secreted from breast tumor cells, but
not normal breast epithelial cells, induced IL-23 protein secretion in dendritic cells via induction of p19 mRNA
expression. Our data further indicate that tumor-secreted PGE2 through EP2 and EP4 receptors enhanced IL-23 p19
gene transcription through binding to the cAMP-response element in the p19 promoter. Blocking PGE2 synthesis by
NS398, a COX2 inhibitor, abrogated the enhancement of p19 expression both in vitro and in vivo. Furthermore,
blocking protein kinase A (PKA) by H89 completely abrogated the inductive effects of tumor-conditioned medium and
PGE2 on p19 transcription, whereas the cAMP active analog, Forskolin, mimics the PGE2 effect. Taken together, our
results indicate that tumor-secreted PGE2 induces IL-23, but not IL-12, production in the tumor microenvironment,
leading to Th17 cell expansion. This inductive effect of PGE2 on IL-23 p19 transcription is mediated through
cAMP/PKA signaling transduction pathway.
~15 Citings
362. Corticosteroid production in H295R cells during exposure to 3 endocrine disrupters analyzed with LC-
MS/MS
By Winther Christina S; Nielsen Frederik K; Hansen Martin; Styrishave Bjarne
From International journal of toxicology (2013), 32(3), 219-27, Language: English, Database: MEDLINE
The adrenocortical human cell line H295R is a valuable tool for screening endocrine disrupting compounds. In
general, previous research focus has been on the production of the 2 sex steroids, 17β-estradiol and testosterone, and
less attention has been paid to other important steroid end points in the steroidogenesis with a wide range of
physiological functions, such as the glucocorticoids (corticosterone and cortisol). A newly developed and validated
solid phase extraction (SPE) liquid chromatography-mass spectroscopy (LC-MS/MS) method was used to measure the
production of cortisol and corticosterone in the H295R cell line. The method was applied by studying the effects of 2
model endocrine disrupters, ketoconazole and prochloraz, the pharmaceutical budesonide, and the inducer forskolin
on the steroid production in this cell line. Dose-response curves were obtained for the correlation between hormone
concentrations and the concentration of the individual disruptors. Exposing cells to ketoconazole resulted in a
decrease in cortisol and corticosterone concentrations in a dose-dependent manner with EC50 values of 0.24 and 0.40
µmol/L, respectively. The same applied for cells exposed to prochloraz with EC50 values of 0.06 and 0.09 µmol/L for
cortisol and corticosterone, respectively. Budesonide also inhibited glucocorticoid secretion. The EC50 value for
cortisol was 19.50 µmol/L, whereas the EC50 value for corticosterone was 71.42 µmol/L. Forskolin induced the
secretion of both cortisol (EC50 = 4.09 µmol/L) and corticosterone (EC50 = 0.28 µmol/L). The results obtained
demonstrated the validity of the method. Based on these findings, quality criteria for the production of these steroids in
this cell line were suggested.
~0 Citings
363. TSH induces metallothionein 1 in thyrocytes via Gq/11- and PKC-dependent signaling
By Back Christer M; Stohr Stefanie; Schafer Eva A M; Biebermann Heike; Boekhoff Ingrid; Breit Andreas; Gudermann
Thomas; Buch Thomas R H
From Journal of molecular endocrinology (2013), 51(1), 79-90, Language: English, Database: MEDLINE
Metallothioneins (MTs) are cytoprotective proteins acting as scavengers of toxic metal ions or reactive oxygen species.
MTs are upregulated in follicular thyroid carcinoma and are regarded as a marker of thyroid stress in Graves' disease.
However, the mechanism of MT regulation in thyrocytes is still elusive. In other cellular systems, cAMP-, calcium-, or
protein kinase C (PKC)-dependent signaling cascades have been shown to induce MT expression. Of note, all of
these three pathways are activated following the stimulation of the TSH receptor (TSHR). Thus, we hypothesized that
TSH represents a key regulator of MT expression in thyrocytes. In fact, TSHR stimulation induced expression of MT
isoform 1X (MT1X) in human follicular carcinoma cells. In these cells, Induction of MT1X expression critically relied on
intact Gq/11 signaling of the TSHR and was blocked by chelation of intracellular calcium and inhibition of PKC. TSHR-
independent stimulation of cAMP formation by treating cells with forskolin also led to an upregulation of MT1X, which
was completely dependent on PKA. However, inhibition of PKA did not affect the regulation of MT1X by TSH. As in
follicular thyroid carcinoma cells, TSH also induced MT1 protein in primary human thyrocytes, which was PKC
dependent as well. In summary, these findings indicate that TSH stimulation induces MT1X expression via Gq/11 and
PKC, whereas cAMP-PKA signaling does not play a predominant role. To date, little has been known regarding cAMP-
independent effects of TSHR signaling. Our findings extend the knowledge about the PKC-mediated functions of the
TSHR.
~0 Citings
364. CREB-regulated transcription co-activator family stimulates promoter II-driven aromatase expression in
preadipocytes
By Samarajeewa Nirukshi U; Docanto Maria M; Simpson Evan R; Brown Kristy A
From Hormones & cancer (2013), 4(4), 233-41, Language: English, Database: MEDLINE
The dramatically increased prevalence of breast cancer after menopause is of great concern and is correlated with
elevated local levels of estrogens. This is mainly due to an increase in aromatase expression driven by its proximal
promoter II (PII). We have previously demonstrated that the CREB co-activator CRTC2 binds directly to PII and
stimulates its activity via mechanisms involving LKB1-AMPK in response to prostaglandin E(2) (PGE(2)). There are
three members of the CRTC family (CRTC1-3) and this study aimed to characterize the role of other CRTCs in the
activation of aromatase PII. The expression and subcellular localization of CRTCs were examined in preadipocytes
using qPCR and immunofluorescence. Under basal conditions, CRTC1 expression was the lowest, whereas CRTC3
transcripts were present at higher levels. Basally, CRTC2 and CRTC3 were mainly cytoplasmic and PGE(2) caused
their nuclear translocation. Reporter assays and chromatin immunoprecipitation (ChIP) were performed to assess the
effect of CRTCs on PII activity and binding. Basal PII activity was significantly increased with all CRTCs. Forskolin
(FSK)/phorbol 12-myristate 13-acetate (PMA), to mimic PGE(2), resulted in a further significant increase in PII activity
with all CRTCs, with CRTC2 and CRTC3 having greater effects. This was consistent with ChIP data showing an
increased binding of CRTCs to PII with FSK/PMA. Moreover, gene silencing of CRTC2 and CRTC3 significantly
reduced the FSK/PMA-mediated stimulation of aromatase activity. Interestingly, CRTCs acted cooperatively with
CREB1 to increase PII activity, and both CREs were found to be essential for the maximal induction of PII activity by
CRTCs. Phosphorylation of CRTC2 at its AMPK target site, Ser 171, dictated its subcellular localization, and the
activation of aromatase PII in preadipocytes. In conclusion, this study demonstrates that aromatase regulation in
primary human breast preadipocytes involves more than one CRTC.
~3 Citings
365. The tumor suppressor RASSF10 is upregulated upon contact inhibition and frequently epigenetically
silenced in cancer
By Richter A M; Walesch S K; Wurl P; Taubert H; Dammann R H
From Oncogenesis (2012), 1e18, Language: English, Database: MEDLINE
The Ras association domain family (RASSF) comprises a group of tumor suppressors that are frequently epigenetically
inactivated in various tumor entities and linked to apoptosis, cell cycle control and microtubule stability. In this work,
we concentrated on the newly identified putative tumor suppressor RASSF10. Methylation analysis reveals RASSF10
promoter hypermethylation in lung cancer, head and neck (HN) cancer, sarcoma and pancreatic cancer. An increase
in RASSF10 methylation from normal tissues, primary tumors to cancer cell lines was observed. Methylation was
reversed by 5-aza-2'-deoxycytidine treatment leading to reexpression of RASSF10. We further show that
overexpression of RASSF10 suppresses colony formation in cancer cell lines. In addition, RASSF10 is upregulated by
cell-cell contact and regulated on promoter level as well as endogenously by forskolin, protein kinase A (PKA) and
activator Protein 1 (AP-1), linking RASSF10 to the cAMP signaling pathway. Knockdown of the AP-1 member JunD
interfered with contact inhibition induced RASSF10 expression. In summary, we found RASSF10 to be epigenetically
inactivated by hypermethylation of its CpG island promoter in lung, HN, sarcoma and pancreatic cancer. Furthermore,
our novel findings suggest that tumor suppressor RASSF10 is upregulated by PKA and JunD signaling upon contact
inhibition and that RASSF10 suppresses growth of cancer cells.
~4 Citings
366. MTA3 regulates CGB5 and Snail genes in trophoblast

By Chen Ying; Miyazaki Jun; Nishizawa Haruki; Kurahashi Hiroki; Leach Richard; Wang Kai
MEDLINE
Secreted by the placental trophoblast, human chorionic gonadotropin (hCG) is an important hormone during pregnancy
and is required for the maintenance of pregnancy. Previous studies have shown that dys-regulation of hCG expression
is associated with preeclampsia. However, the exact relationship between altered hCG levels and development of
preeclampsia is unknown. Metastasis associated protein 3 (MTA3), a chromatin remodeling protein, is abundantly
expressed in the placental trophoblasts, but its function is unknown. In breast cancer, MTA3 has been shown to
repress the expression of Snail and cell migration. However, whether MTA3 acts similarly in the trophoblast has not
been investigated. In the present study, we examined the role of MTA3 in regulating the hCG β-subunit gene (gene
name: CGB5) and Snail expression in the trophoblast cell line, BeWo, as well as its relevance to the high hCG
expression levels seen in preeclampsia. First, we investigated MTA3 expression in preeclamptic placenta as
compared to normal control placenta via gene expression microarray and qRT-PCR and found that MTA3 was
significantly down-regulated, whereas both CGB5 and Snail were up-regulated in preeclamptic placenta. Secondly, we
knocked down MTA3 gene in trophoblast cell line BeWo and found Snail and hCG were both up-regulated, suggesting
that MTA3 represses Snail and hCG gene expression in trophoblasts. Next, we cloned the CGB5 and Snail promoters
into the pGL3-basic vector individually and found that silencing of MTA3 by siRNA resulted in an increase of both
CGB5 and Snail promoter activities. To confirm that this MTA3 inhibition is a direct effect, we performed a chromatin
immune-precipitation (ChIP) assay and found that MTA3 occupied the proximal promoter regions of both Snail and
hCG within BeWo cells. Furthermore, we examined MTA3 expression in placental trophoblast by
immunohistochemistry and found that MTA3 expression was higher in villous cytotrophoblasts versus
syncytiotrophoblasts, which supports an inverse association of MTA3 with hCG expression. Lastly, using the well-
characterized trophoblast fusion model, we examined MTA3 and hCG levels in forskolin-treated BeWo cells and found
that MTA3 down-regulation was accompanied by an up-regulation of hCG. These data further suggest that MTA3 is
repressing placental hCG expression. In summary, MTA3 plays a critical role in repressing hCG and Snail in placenta
trophoblast and its deregulation is associated with preeclampsia.
~3 Citings
367. Discovery, molecular and pharmacological characterization of GSA-10, a novel small-molecule positive
modulator of Smoothened
By Gorojankina Tatiana; Hoch Lucile; Faure Helene; Roudaut Hermine; Traiffort Elisabeth; Schoenfelder Angele;
Girard Nicolas; Mann Andre; Manetti Fabrizio; Solinas Antonio; et al
From Molecular pharmacology (2013), 83(5), 1020-9, Language: English, Database: MEDLINE
Activation of the Smoothened (Smo) receptor mediates Hedgehog (Hh) signaling. Hh inhibitors are in clinical trials for
cancer, and small-molecule Smo agonists may have therapeutic interests in regenerative medicine. Here, we have
generated and validated a pharmacophoric model for Smo agonists and used this model for the virtual screening of a
library of commercially available compounds. Among the 20 top-scoring ligands, we have identified and characterized
a novel quinolinecarboxamide derivative, propyl 4-(1-hexyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carboxamido)
benzoate, (GSA-10), as a Smo agonist. GSA-10 fits to the agonist pharmacophoric model with two hydrogen bond
acceptor groups and four hydrophobic regions. Using pharmacological, biochemical, and molecular approaches, we
provide compelling evidence that GSA-10 acts at Smo to promote the differentiation of multipotent mesenchymal
progenitor cells into osteoblasts. However, this molecule does not display the hallmarks of reference Smo agonists.
Remarkably, GSA-10 does not recognize the classic bodipy-cyclopamine binding site. Its effect on cell differentiation is
inhibited by Smo antagonists, such as MRT-83, SANT-1, LDE225, and M25 in the nanomolar range, by GDC-0449 in
the micromolar range, but not by cyclopamine and CUR61414. Thus, GSA-10 allows the pharmacological
characterization of a novel Smo active site, which is notably not targeted to the primary cilium and strongly potentiated
by forskolin and cholera toxin. GSA-10 belongs to a new class of Smo agonists and will be helpful for dissecting Hh
mechanism of action, with important implications in physiology and in therapy.
~5 Citings
368. Protein kinase A (PKA) pathway is functionally linked to androgen receptor (AR) in the progression of
prostate cancer
By Sarwar Martuza; Sandberg Sabina; Abrahamsson Per-Anders; Persson Jenny L
From Urologic oncology (2014), 32(1), 25.e1-12, Language: English, Database: MEDLINE
OBJECTIVES: In the present study, we investigated whether the cyclic adenosine monophosphate (cAMP)-activated
protein kinase A (PKA) pathway may regulate the expression of AR and prostate-specific antigen (PSA) and whether
there is a correlation between the expression of cAMP/PKA-associated genes and androgen receptor (AR) in patients
with prostate cancer (CaP). MATERIALS AND METHODS: The functional studies were performed in LNCaP and PC3
cell lines. Data on the mRNA expression of sets of genes in human clinical samples, including prostate tissues from
organ donors, prostate primary cancer, and metastatic cancer, were extracted from the National Center for
Biotechnology Informations Gene Expression Omnibus (GEO) database. Statistical tests were applied. RESULTS:
We showed that elevated levels of cAMP/PKA pathways induced an increased expression of AR and PSA proteins in
LNCaP cells in the absence of androgen. A cAMP-associated phosphodiesterase-4 (PDE4) inhibitor, rolipram induced
an up-regulation in AR expression, whereas a cAMP enhancer, forskolin increased PSA level without affecting AR
expression. Forskolin treatment increased the level of PKA R1α in LNCaP cells, but remarkably inhibited R1α
expression in aggressive PC3 cells. In patients with CaP, we found that the expression of genes encoding R1α and
phosphodiesterase-4B was statistically significantly lower in the metastatic specimens than that in the primary CaP
specimens or in the normal prostate tissues (P<0.01) and was reversely correlated with AR expression. Conversely,
AR and PRKAR2B mRNA expressions were significantly higher in metastatic lesions than those in the primary CaP
specimens or in the normal prostate tissues (P<0.01). CONCLUSION: Our study revealed a novel mechanism to
precisely define the functional and clinical interrelationship between the cAMP/PKA pathway and AR signaling in the
development of androgen-independent growth of CaPs and metastasis progression.
~8 Citings
369. Protection against radiation-induced damage of 6-propyl-2-thiouracil (PTU) in thyroid cells

By Perona Marina; Dagrosa Maria A; Pagotto Romina; Casal Mariana; Pignataro Omar P; Pisarev Mario A; Juvenal
Guillermo J
From Radiation research (2013), 179(3), 352-60, Language: English, Database: MEDLINE
Many epidemiologic studies have shown that the exposure to high external radiation doses increases thyroid
neoplastic frequency, especially when given during childhood or adolescence. The use of radioprotective drugs may
decrease the damage caused by radiation therapy and therefore could be useful to prevent the development of thyroid
tumors. The aim of this study was to investigate the possible application of 6-propyl-2-thiouracil (PTU) as a
radioprotector in the thyroid gland. Rat thyroid epithelial cells (FRTL-5) were exposed to different doses of γ irradiation
with or without the addition of PTU, methimazole (MMI), reduced glutathione (GSH) and perchlorate (KClO4).
Radiation response was analyzed by clonogenic survival assay. Cyclic AMP (cAMP) levels were measured by
radioimmunoassay (RIA). Apoptosis was quantified by nuclear cell morphology and caspase 3 activity assays.
Intracellular reactive oxygen species (ROS) levels were measured using the fluorescent dye 2',7'-dichlorofluorescein-
diacetate. Catalase, superoxide dismutase and glutathione peroxidase activities were also determined. Pretreatment
with PTU, MMI and GSH prior to irradiation significantly increased the surviving cell fraction (SF) at 2 Gy (P < 0.05),
while no effect was observed with KClO4. An increase in extracellular levels of cAMP was found only in PTU treated
cells in a dose and time-dependent manner. Cells incubated with agents that stimulate cAMP (forskolin and dibutyril
cAMP) mimicked the effect of PTU on SF. Moreover, pretreatment with the inhibitor of protein kinase A, H-89,
abolished the radioprotective effect of PTU. PTU treatment diminished radiation-induced apoptosis and protected cells
against radiation-induced ROS elevation and suppression of the antioxidant enzyme's activity. PTU was found to
radioprotect normal thyroid cells through cAMP elevation and reduction in both apoptosis and radiation-induced
oxidative stress damage.
~0 Citings
370. Effect of polyphenols on production of steroid hormones from human adrenocortical NCI-H295R cells
By Hasegawa Eri; Nakagawa Saori; Sato Momoe; Tachikawa Eiichi; Yamato Susumu
From Biological & pharmaceutical bulletin (2013), 36(2), 228-37, Language: English, Database: MEDLINE
Modulating steroid hormone levels is a curative and preventive measure for Cushing's syndrome, aldosteronism, and
various stress-triggered symptoms. Polyphenols have been reported to inhibit steroidogenic enzymes such as 3β-
hydroxysteroid dehydrogenase (3β-HSD) and aromatase. However, evidence for their inhibitory effects is fragmentary
because it has been determined in studies with small groups of steroid hormones. To investigate the effects of
steroids on complete steroidogenic pathways, comprehensive analysis of steroid hormones is necessary. Here we
cultured forskolin-stimulated NCI-H295R, a human adrenocortical carcinoma cell line, in the presence of a polyphenol
and employed GC-MS to simultaneously determine the levels of nine steroid hormones (pregnenolone, progesterone,
deoxycorticosterone, aldosterone, 17α-hydroxyprogesterone, dehydroepiandrosterone, androstenedione, testosterone,
and estradiol) in cell culture supernatant. We found that daidzein, genistein, apigenin, hesperetin, naringenin, and
eriodictyol significantly reduced deoxycorticosterone and androstenedione levels (p<0.05), suggesting inhibition of 3β-
HSD by these polyphenols. Apigenin was more potent than other polyphenols in increasing the levels of pregnenolone
and 17α-hydroxyprogesterone, suggesting that it inhibits cytochrome P450 (CYP) 17 and CYP21, as well as 3β-HSD.
Real-time reverse transcription polymerase chain reaction showed that apigenin significantly downregulated the
expression levels of 3β-HSD, CYP17, and CYP21 mRNA (p<0.05). This is the first study to demonstrate the inhibitory
effects of apigenin on CYP17 and CYP21.
~0 Citings
371. Aglycone of Rh4 inhibits melanin synthesis in B16 melanoma cells: possible involvement of the protein
kinase A pathway
By Jeong Yun-Mi; Oh Won Keun; Tran Tien Lam; Kim Wang-Kyun; Sung Sang Hyun; Bae Kyeol; Lee Suman; Sung
Jong-Hyuk
From Bioscience, biotechnology, and biochemistry (2013), 77(1), 119-25, Language: English, Database: MEDLINE
To our knowledge, there is no report that directly shows an inhibitory effect of ginsenoside on melanin synthesis in B16
melanoma cells. Hence, we investigated whether the aglycone of Rh(4) (A-Rh4) inhibits melanin synthesis in B16
melanoma cells, and determined the mechanism of melanin inhibition. We isolated 12 ginsenoside compounds from
leaves of Panax ginseng and tested them in B16 melanoma cells. It significantly reduced melanin content and
tyrosinase activity under alpha-melanocyte stimulating hormone- and forskolin-stimulated conditions. It significantly
reduced the cyclic AMP (cAMP) level in B16 melanoma cells, and this might be responsible for the regulation down of
MITF and tyrosinase. Phosphorylation of a downstream molecule, a cAMP response-element binding protein, was
significantly decreased according to Western blotting and immunofluorescence assay. These data suggest that A-Rh4
has an anti-melanogenic effect via the protein kinase A pathway.
~2 Citings
372. Prostaglandin E2 induces stromal cell-derived factor-1 expression in prostate stromal cells by activating
protein kinase A and transcription factor Sp1
By Peng Yanfei; Shi Jiandang; Du Xiaoling; Wang Liang; Klocker Helmut; Mo Linjian; Mo Zengnan; Zhang Ju
From The international journal of biochemistry & cell biology (2013), 45(3), 521-30, Language: English, Database:
MEDLINE
Recent reports indicate prostaglandin E2 (PGE2) can modulate tumor environment and promote angiogenesis through
induction of stromal cell-derived factor 1 (SDF-1) production. We investigated the mechanism of PGE2-induced SDF-1
regulation in human prostate stromal cell and analyzed the effects in a stromal-epithelial interaction model. PGE2
stimulation increased SDF-1 expression in the prostate stromal cell lines WPMY-1 and NAF. We revealed signaling
through the PGE2 receptor EP3 and activation of protein kinase A (PKA) are required. The EP3 agonist sulprostone
and the cAMP analog forskolin mimicked and the EP3 siRNA, antagonist L798106 and the PKA inhibitor H89
abrogated the effect of PGE2 on SDF-1 expression. SDF-1 promoter truncation experiments demonstrated a 254 bp
(from nt -219 to nt +34) SDF-1 proximal promoter fragment containing 5 putative transcription factor Sp1 binding motifs
is sufficient for PGE2 induction. CHIP assays confirmed binding and PGE2 induced recruitment of Sp1 to the SDF-1
promoter. Sp1 motif mutation identified Sp1 motifs -140/-133 and -9/+1 as the crucial elements responsible for PGE2
induction. Moreover, SDF-1 was up- or down-regulated by Sp1 over-expression or knock-down. We also demonstrate
stimulation of migration of prostate cancer cell lines PC3 and DU145 with conditioned media collected from WPMY-1 or
NAF cells stimulated with PGE2 and blockade of enhanced migration by a SDF-1 neutralizing antibody. In conclusion,
we provide evidence for a paracrine prostate stromal-epithelial interaction induced by upregulation of expression of
SDF-1 by PGE2. Our research provides new insights into the mechanism promoting metastasis of prostate carcinoma
via stromal-epithelial interaction.
~1 Citing
373. RCAN1 is an important mediator of glucocorticoid-induced apoptosis in human leukemic cells

By Nagao Kazuaki; Iwai Yujiro; Miyashita Toshiyuki
Glucocorticoid (GC) is a major therapeutic agent for the treatment of leukemia because of its ability to induce apoptosis
in lymphoid cells. The mechanism causing apoptosis, however, is still controversial. Since the glucocorticoid receptor
is a transcription factor, some of its target genes are expected to be implicated in apoptosis. In this study, using a GC-
sensitive human pre-B leukemia cell line, Nalm-6, the FK506 binding protein 51 (FKBP5) and regulator of calcineurin 1
(RCAN1) genes were disrupted by homologous recombination, since the expression of both is up-regulated by GC in
GC-sensitive but not in GC-resistant leukemic cell lines. While the disruption of FKBP5 had a marginal effect on GC-
induced apoptosis, that of RCAN1 resulted in marked resistance to GC. In addition, overexpression of RCAN1
rendered cells more sensitive to DEX. In RCAN1-disrupted cells, levels of some pro-apoptotic and anti-apoptotic Bcl-2
family proteins were decreased and increased, respectively. Finally, phosphorylation of cAMP-response element
binding protein (CREB) and up-regulation of CREB target genes by GC were inhibited by RCAN1 disruption, and
treatment with a cAMP-inducing agent, forskolin, restored the sensitivity to GC in RCAN1-disrupted Nalm-6 cells.
These findings suggest that up-regulation of RCAN1 expression followed by activation of the CREB pathway is
required in GC-induced apoptosis.
~3 Citings
374. Inhibition of CIP2A determines erlotinib-induced apoptosis in hepatocellular carcinoma

By Yu Hui-Chuan; Chen Hui-Ju; Chang Ya-Ling; Liu Chun-Yu; Shiau Chung-Wai; Cheng Ann-Lii; Chen Kuen-Feng
Erlotinib is a small-molecular inhibitor of epidermal growth factor receptor (EGFR). Here, we identify that cancerous
inhibitor of protein phosphatase 2A (CIP2A) is a major determinant mediating erlotinib-induced apoptosis in
hepatocellular carcinoma (HCC). Erlotinib showed differential effects on apoptosis in 4 human HCC cell lines.
Erlotinib induced significant apoptosis in Hep3B and PLC5 cell lines; however, Huh-7 and HA59T cell lines showed
resistance to erlotinib-induced apoptosis at all tested doses. Down-regulation of CIP2A, a cellular inhibitor of protein
phosphatase 2A (PP2A), mediated the apoptotic effect of erlotinib in HCC. Erlotinib inhibited CIP2A in a dose- and
time-dependent manner in all sensitive HCC cells whereas no alterations in CIP2A were found in resistant cells.
Overexpression of CIP2A upregulated phospho-Akt and protected Hep3B cells from erlotinib-induced apoptosis. In
addition, silencing CIP2A by siRNA restored the effects of erlotinib in Huh-7 cells. Moreover, adding okadaic acid, a
PP2A inhibitor, abolished the effects of erlotinib on apoptosis in Hep3B cells; and forskolin, a PP2A agonist enhanced
the effect of erlotinib in resistant HA59T cells. Combining Akt inhibitor MK-2206 with erlotinib restored the sensitivity of
HA59T cells to erlotinib. Furthermore, in vivo xenograft data showed that erlotinib inhibited the growth of PLC5 tumor
but had no effect on Huh-7 tumor. Erlotinib downregulated CIP2A and upregulated PP2A activity in PLC5 tumors, but
not in Huh-7 tumors. In conclusion, inhibition of CIP2A determines the effects of erlotinib on apoptosis in HCC. CIP2A
may be useful as a therapeutic biomarker for predicting clinical response to erlotinib in HCC treatment.
~9 Citings
375. A(3) adenosine receptor mediates apoptosis in 5637 human bladder cancer cells by G(q) protein/PKC-
dependent AIF upregulation
By Kanno Takeshi; Gotoh Akinobu; Fujita Yumiko; Nakano Takashi; Nishizaki Tomoyuki
From Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and
pharmacology (2012), 30(5), 1159-68, Language: English, Database: MEDLINE
BACKGROUND/AIMS: A(3) adenosine receptor mediates apoptosis in a variety of cancer cells via diverse signaling
pathways. The present study was conducted to assess A(3) adenosine receptor-mediated apoptosis in human bladder
cancer cell lines and to understand the underlying mechanism. METHODS: Human bladder cancer cell lines such as
253J, 5637, KK-47, TCCSUP, T24, and UMUC-3 cells were cultured. The siRNA to silence the A(3) adenosine
receptor-targeted gene was constructed and transfected into cells. MTT assay, TUNEL staining, Western blotting, and
real-time RT-PCR were carried out. RESULTS: For all the investigated cell types adenosine induced apoptosis in a
concentration (0.01-10 mM)- and treatment time (24-48 h)-dependent manner. Adenosine-induced 5637 cell death
was significantly inhibited by the A(3) adenosine receptor inhibitor MRS1191 or knocking-down A(3) adenosine
receptor, and the A(3) adenosine receptor agonist 2-Cl-IB-MECA mimicked the adenosine effect. The adenosine effect
was prevented by GF109203X, an inhibitor of protein kinase C (PKC), but it was not affected by forskolin, an activator
of adenylate cyclase. Adenosine-induced 5637 cell death, alternatively, was not inhibited by the pan-caspase inhibitor
Z-VAD. Adenosine upregulated expression of apoptosis-inducing factor (AIF), that is suppressed by knocking-down
A(3) adenosine receptor, and accumulated AIF in the nucleus. CONCLUSION: The results of the present study show
that adenosine induces 5637 cell apoptosis by upregulating AIF expression via an A(3) adenosine receptor-mediated
G(q) protein/PKC pathway.
~1 Citing
376. 5-aza-2'-deoxycytidine has minor effects on differentiation in human thyroid cancer cell lines, but
modulates genes that are involved in adaptation in vitro
By Dom Genevieve; Galdo Vanessa Chico; Tarabichi Maxime; Tomas Gil; Hebrant Aline; Andry Guy; De Martelar
Viviane; Libert Frederick; Leteurtre Emmanuelle; Dumont Jacques E; et al
From Thyroid : official journal of the American Thyroid Association (2013), 23(3), 317-28, Language: English,
Database: MEDLINE
BACKGROUND: In thyroid cancer, the lack of response to specific treatment, for example, radioactive iodine, can be
caused by a loss of differentiation characteristics of tumor cells. It is hypothesized that this loss is due to epigenetic
modifications. Therefore, drugs releasing epigenetic repression have been proposed to reverse this silencing.
METHODS: We investigated which genes were reinduced in dedifferentiated human thyroid cancer cell lines when
treated with the demethylating agent 5-aza-2'-deoxycytidine (5-AzadC) and the histone deacetylase inhibitors
trichostatin A (TSA) and suberoylanilide hydroxamic acid, by using reverse transcriptase-polymerase chain reaction
and microarrays. These results were compared to the expression patterns in in vitro human differentiated thyrocytes
and in in vivo dedifferentiated thyroid cancers. In addition, the effects of 5-AzadC on DNA quantities and cell viability
were investigated. RESULTS: Among the canonical thyroid differentiation markers, most were not, or only to a minor
extent, re-expressed by 5-AzadC, whether or not combined with TSA or forskolin, an inducer of differentiation in normal
thyrocytes. Furthermore, 5-AzadC-modulated overall mRNA expression profiles showed only few commonly regulated
genes compared to differentiated cultured primary thyrocytes. In addition, most of the commonly strongly 5-AzadC-
induced genes in cell lines were either not regulated or upregulated in anaplastic thyroid carcinomas. Further analysis
of which genes were induced by 5-AzadC showed that they were involved in pathways such as apoptosis, antigen
presentation, defense response, and cell migration. A number of these genes had similar expression responses in 5-
AzadC-treated nonthyroid cell lines. CONCLUSIONS: Our results suggest that 5-AzadC is not a strong inducer of
differentiation in thyroid cancer cell lines. Under the studied conditions and with the model used, 5-AzadC treatment
does not appear to be a potential redifferentiation treatment for dedifferentiated thyroid cancer. However, this may
reflect primarily the inadequacy of the model rather than that of the treatment. Moreover, the observation that 5-AzadC
negatively affected cell viability in cell lines could still suggest a therapeutic opportunity. Some of the genes that were
modulated by 5-AzadC were also induced in nonthyroid cancer cell lines, which might be explained by an epigenetic
modification resulting in the adaptation of the cell lines to their culture conditions.
~2 Citings
377. Protein kinase A activation inhibits oncogenic Sonic hedgehog signalling and suppresses basal cell
carcinoma of the skin
By Makinodan Eri; Marneros Alexander G
From Experimental dermatology (2012), 21(11), 847-52, Language: English, Database: MEDLINE
Basal cell carcinoma of the skin (BCC) is caused by constitutive activation of the Sonic hedgehog (Shh) pathway,
mainly through mutations either in the Shh receptor Patched (PTCH) or in its co-receptor Smoothened (Smo).
Inhibitors of this pathway that are currently in clinical trials inhibit Smo. However, mutations in Smo can result in
resistance to these inhibitors. To target most BCCs and avoid acquired resistance because of Smo mutations,
inhibiting the Shh-pathway downstream of Smo is critical. Attractive downstream targets would be at the level of Gli
proteins, the transcriptional activators of this pathway in BCCs. Previously it has been shown that Gli1 and Gli2, when
phosphorylated by protein kinase A (PKA), are targeted for proteosomal degradation. Here we show that PKA
activation via the cAMP agonist forskolin is sufficient to completely abolish oncogenic Smo activity in vitro. In an
inducible BCC mouse model due to a Smo mutation that confers resistance to current Smo inhibitors, topical forskolin
treatment significantly reduced Gli1 mRNA levels and resulted in strongly suppressed BCC tumor growth. Our data
show that forskolin inhibits the growth of even those BCCs that are resistant to Smo inhibitors and provide a proof-of-
principle framework for the development of topically applied human skin-permeable novel pharmacologic inhibitors of
oncogenic Shh-signaling through PKA activation.
~4 Citings
378. Suppression of melanin synthesis by the phenolic constituents of sappanwood (Caesalpinia sappan)
By Mitani Kaoru; Takano Fumihide; Kawabata Tetsuro; Allam Ahmed E; Ota Mayumi; Takahashi Tomoya; Yahagi
Nobuo; Sakurada Chikai; Fushiya Shinji; Ohta Tomihisa
From Planta medica (2013), 79(1), 37-44, Language: English, Database: MEDLINE
Sappanwood (Caesalpinia sappan Linn.) is used as an herbal medicine. It is sometimes used to treat skin damage or
as a facial cleanser. In the present study, the methanol (MeOH) extract of sappanwood was found to inhibit melanin
synthesis in cultured human melanoma HMV-II cells stimulated with forskolin, and six active compounds (1-5 and 7)
were isolated from the extract along with a non-active compound (6). Compounds 2-7 were identified as
sappanchalcone (2), 3'-deoxy-4-O-methylsappanol (3), brazilein, (4), brazilin (5), sappanol (6), and 4-O-
methylsappanol (7). Compound 1 was a new compound, and its structure was determined to be (6aS,11bR)-7,11b-
dihydro-6H-indeno[2,1-c]chromene-3,6a,10,11-tetrol by spectroscopic analyses. Among the six active compounds,
brazilin (5) (EC50: 3.0 ± 0.5 µM) and 4-O-methylsappanol (7) (EC50: 4.6 ± 0.7 µM) strongly suppressed melanin
synthesis in HMV-II cells. Bioactive compounds showed moderate cytotoxicities against HMV-II cells with IC50 values
of 83.1 ± 4.0 µM (for 2), 72.0 µM ± 2.4 (for 3), 33.8 ± 1.1 µM (for 4), 18.4 ± 0.8 µM (for 5), and 20.2 ± 0.8 (for 7),
respectively. Brazilin (5) selectively suppressed the expression of mRNAs for tyrosinase-related protein (TYRP) 2 and
tyrosinase but did not influence the expression of TYRP1. These results suggest that brazilin (5) is a new class of
melanin inhibitor and that sappanwood could be used as a cosmetic material.
~3 Citings
379. Identification of sumoylation sites in CCDC6, the first identified RET partner gene in papillary thyroid
carcinoma, uncovers a mode of regulating CCDC6 function on CREB1 transcriptional activity
By Luise Chiara; Merolla Francesco; Leone Vincenza; Paladino Simona; Sarnataro Daniela; Fusco Alfredo; Celetti
Angela
CCDC6 was originally identified in chimeric genes as caused by chromosomal translocation involving the RET
protooncogene in some thyroid tumors. Recognised as a 65 kDa pro-apoptotic phosphoprotein, CCDC6 has been
enrolled as an ATM substrate that contribute to protect genome integrity by modulating PP4c activity in response to
genotoxic stress. Recently, CCDC6 has been identified as a repressor of CREB1-dependent transcription.
Sumoylation has emerged as an important mechanism in transcriptional control. Here, we report the identification and
characterization of three sites of sumoylation in CCDC6 (K74, K266 and K424) which are highly conserved in
vertebrates. We demonstrate that the post-translational modifications by SUMO2 constrain most of the CCDC6 protein
in the cytosol and affect its functional interaction with CREB1 with a decrease of CCDC6 repressive function on
CREB1 transcriptional activity. Indeed, the impairment of functional outcome of sumoylated CCDC6 is obtained
knocking down all three the sumoylation sites. Interestingly, in thyroid cells the SUMO2-mediated CCDC6 post-
translational modifications are induced by Forskolin, a cAMP analog. Signal transduction via the cAMP pathway is
known to be ubiquitous and represents a major line of communication between many organisms and their environment.
We believe that CCDC6 could be an important player in the dynamics of cAMP signaling by fine regulating CREB1
transcriptional activity in normal and transformed thyroid cells.
~2 Citings
380. Pigment-independent cAMP-mediated epidermal thickening protects against cutaneous UV injury by

keratinocyte proliferation
By Scott Timothy L; Christian Perry A; Kesler Melissa V; Donohue Kevin M; Shelton Brent; Wakamatsu Kazumasa; Ito
Shosuke; D'Orazio John
From Experimental dermatology (2012), 21(10), 771-7, Language: English, Database: MEDLINE
The epidermis increases pigmentation and epidermal thickness in response to ultraviolet exposure to protect against
UV-associated carcinogenesis; however, the contribution of epidermal thickness has been debated. In a humanized
skin mouse model that maintains interfollicular epidermal melanocytes, we found that forskolin, a small molecule that
directly activates adenylyl cyclase and promotes cAMP generation, up-regulated epidermal eumelanin accumulation in
fair-skinned melanocortin-1-receptor (Mc1r)-defective animals. Forskolin-induced pigmentation was associated with a
reproducible expansion of epidermal thickness irrespective of melanization or the presence of epidermal melanocytes.
Rather, forskolin-enhanced epidermal thickening was mediated through increased keratinocyte proliferation, indirectly
through secreted factor(s) from cutaneous fibroblasts. We identified keratinocyte growth factor (Kgf) as a forskolin-
induced fibroblast-derived cytokine that promoted keratinocyte proliferation, as forskolin induced Kgf expression both in
the skin and in primary fibroblasts. Lastly, we found that even in the absence of pigmentation, forskolin-induced
epidermal thickening significantly diminished the amount of UV-A and UV-B that passed through whole skin and
reduced the amount of UV-B-associated epidermal sunburn cells. These findings suggest the possibility of
pharmacologic-induced epidermal thickening as a novel UV-protective therapeutic intervention, particularly for
individuals with defects in pigmentation and adaptive melanization.
~5 Citings
381. Functional status and relationships of melanocortin 1 receptor signaling to the cAMP and extracellular
signal-regulated protein kinases 1 and 2 pathways in human melanoma cells
By Herraiz Cecilia; Journe Fabrice; Ghanem Ghanem; Jimenez-Cervantes Celia; Garcia-Borron Jose C
MEDLINE
Melanocortin 1 receptor (MC1R), a major determinant of skin phototype frequently mutated in melanoma, is a Gs
protein-coupled receptor that regulates pigment production in melanocytes. MC1R stimulation activates cAMP
synthesis and the extracellular signal-regulated (ERK) ERK1 and ERK2. In human melanocytes, ERK activation by
MC1R relies on cAMP-independent transactivation of the c-KIT receptor. Thus MC1R functional coupling to the cAMP
and ERK pathways may involve different structural requirements giving raise to biased effects of skin cancer-
associated mutations. We evaluated the impact of MC1R mutations on ERK activation, cAMP production and agonist
binding. We found that MC1R mutations impair cAMP production much more often than ERK activation, suggesting
less stringent requirements for functional coupling to the ERK pathway. We examined the crosstalk of the cAMP and
ERK pathways in HBL human melanoma cells (wild-type for MC1R, NRAS and BRAF). ERK activation by
constitutively active upstream effectors or pharmacological inhibition had little effect on MC1R-stimulated cAMP
synthesis. High cAMP levels were compatible with normal ERK activation but, surprisingly, the adenylyl cyclase
activator forskolin abolished ERK activation by MC1R, most likely by a cAMP-independent mechanism. These results
indicate little crosstalk of the cAMP and ERK pathways in HBL melanoma cells. Finally, we studied cAMP
accumulation in a panel of 22 human melanoma cell lines stimulated with MC1R agonists or forskolin. cAMP synthesis
was often inhibited, even in cells wild-type for MC1R and NRAS. Therefore, the cAMP pathway is more frequently
impaired in melanoma than could be predicted by the MC1R or NRAS genotype.
~7 Citings
382. Phosphodiesterase 11A (PDE11A) gene defects in patients with acth-independent macronodular adrenal
hyperplasia (AIMAH): functional variants may contribute to genetic susceptibility of bilateral adrenal tumors
By Vezzosi Delphine; Libe Rossella; Baudry Camille; Rizk-Rabin Marthe; Horvath Anelia; Levy Isaac; Rene-Corail
Fernande; Ragazzon Bruno; Stratakis Constantine A; Vandecasteele Gregoire; et al
From The Journal of clinical endocrinology and metabolism (2012), 97(11), E2063-9, Language: English, Database:
MEDLINE
CONTEXT: Phosphodiesterases (PDEs) are key regulatory enzymes of intracellular cAMP levels. PDE11A function
has been linked to predisposition to adrenocortical tumors. OBJECTIVE: The aim of the study was to study the
PDE11A gene in a large cohort of patients with ACTH-independent macronodular adrenal hyperplasia (AIMAH) and in
control subjects. DESIGN: The PDE11A entire coding region was sequenced in 46 patients with AIMAH and 192
controls. Two variants found in AIMAH patients were transiently expressed in HEK 293 and adrenocortical H295R
cells for further functional studies. RESULTS: The frequency of all PDE11A variants was significantly higher among
patients with AIMAH (28%) compared to controls (7.2%) (P = 5 × 10(-5)). Transfection of the two PDE11A variants
found in AIMAH patients only (D609N or M878V) showed that cAMP levels were higher, after forskolin stimulation, in
cells transfected with the PDE11A mutants, compared to cells transfected with the wild-type PDE11A in HEK 293 cells
(P < 0.05). Moreover, transfection with mutants PDE11A increased transcriptional activity of a cAMP-response
element reporter construct compared to wild-type PDE11A in HEK 293 cells (P < 0.0004 for D609N and P < 0.003 for
M878V) and in the adrenocortical H295R cells (P < 0.05 for D609N and M878V). In addition, analysis of cAMP levels
in intact living culture cells by fluorescence resonance energy transfer probes showed increased cAMP in forskolin-
treated cells transfected with PDE11A variants compared with wild-type PDE11A (P < 0.05). CONCLUSION: We
conclude that PDE11A genetic variants may increase predisposition to AIMAH.
~11 Citings
383. cAMP-dependent protein kinase is essential for hypoxia-mediated epithelial-mesenchymal transition,

migration, and invasion in lung cancer cells
By Shaikh Dooniya; Zhou Qiyuan; Chen Tianji; Ibe Joyce Christina F; Raj J Usha; Zhou Guofei
Lung cancer is the leading cause of cancer-related death worldwide. Hypoxia is known to increase cancer cell
migration and invasion. We have previously reported that hypoxia induces epithelial-mesenchymal transition (EMT) in
lung cancer cells. However, it is unknown whether hypoxia promotes lung cancer cell migration and invasion via EMT
and whether cyclic AMP (cAMP) dependent protein kinase (PKA) plays a role in this process. We found that hypoxia
increased PKA activity and induced mRNA and protein expression of PKA catalytic subunit α (PKACA), and regulatory
subunits R1A and R1B. Knockdown of HIF-1/2α prevented hypoxia-mediated induction of PKACA mRNA expression
and PKA activity. Inhibition of PKA activity with chemical inhibitors prevented EMT induced by hypoxia and tumor
growth factor β1. However, activation of PKA by forskolin and 8-Br-cAMP did not induce EMT. Furthermore, treatment
with H89 and knockdown of PKACA prevented hypoxia-mediated, EMT, cell migration, and invasion, whereas
overexpression of mouse PKACA rescued hypoxia-mediated migration and invasion in PKACA deficient cancer cells.
Our results suggest that hypoxia enhances PKA activity by upregulating PKA gene expression in a HIF dependent
mechanism and that PKA plays a key role in hypoxia-mediated EMT, migration, and invasion in lung cancer cells.
~16 Citings
384. The essential role of Giα2 in prostate cancer cell migration

By Zhong Miao; Clarke Shineka; Vo BaoHan T; Khan Shafiq A
Cell- and receptor-specific regulation of cell migration by Gi/oα-proteins remains unknown in prostate cancer cells. In
the present study, oxytocin (OXT) receptor was detected at the protein level in total cell lysates from C81 (an
androgen-independent subline of LNCaP), DU145 and PC3 prostate cancer cells, but not in immortalized normal
prostate luminal epithelial cells (RWPE1), and OXT-induced migration of PC3 cells. This effect of OXT has been
shown to be mediated by Gi/oα-dependent signaling. Accordingly, OXT inhibited forskolin-induced luciferase activity in
PC3 cells that were transfected with a luciferase reporter for cyclic AMP activity. Although mRNAs for all three Giα
isoforms were present in PC3 cells, Giα2 was the most abundant isoform that was detected at the protein level.
Pertussis toxin (PTx) inhibited the OXT-induced migration of PC3 cells. Ectopic expression of the PTx-resistant Giα2-
C352G, but not wild-type Giα2, abolished this effect of PTx on OXT-induced cell migration. The Giα2-targeting siRNA
was shown to specifically reduce Giα2 mRNA and protein in prostate cancer cells. The Giα2-targeting siRNA
eliminated OXT-induced migration of PC3 cells. These data suggest that Giα2 plays an important role in the effects of
OXT on PC3 cell migration. The Giα2-targeting siRNA also inhibited EGF-induced migration of PC3 and DU145 cells.
Expression of the siRNA-resistant Giα2, but not wild type Giα2, restored the effects of EGF in PC3 cells transfected
with the Giα2-targeting siRNA. In conclusion, Giα2 plays an essential role in OXT and EGF signaling to induce
prostate cancer cell migration.
~3 Citings
385. Hormonal induction of polo-like kinases (Plks) and impact of Plk2 on cell cycle progression in the rat ovary
By Li Feixue; Jo Misung; Curry Thomas E Jr; Liu Jing
The highly conserved polo-like kinases (Plks) are potent regulators of multiple functions in the cell cycle before and
during mitotic cell division. We investigated the expression pattern of Plk genes and their potential role(s) in the rat
ovary during the periovulatory period. Plk2 and Plk3 were highly induced both in intact ovaries and granulosa cells in
vivo after treatment with the luteinizing hormone (LH) agonist, human chorionic gonadotropin (hCG). In vitro, hCG
stimulated the expression of Plk2 in granulosa cells, but not Plk3. This induction of Plk2 expression was mimicked by
both forskolin and phorbol 12 myristate 13-acetate (PMA). Moreover, Plk2 expression was reduced by inhibitors of
prostaglandin synthesis or the EGF pathway, but not by progesterone receptor antagonist (RU486) treatment. At the
promoter level, mutation of the Sp1 binding sequence abolished the transcriptional activity of the Plk2 gene. ChIP
assays also revealed the interaction of endogenous Sp1 protein in the Plk2 promoter region. Functionally, the over-
expression of Plk2 and Plk3 arrested granulosa cells at the G0/G1 phase of the cell cycle. In contrast, the knockdown
of Plk2 expression in granulosa cells decreased the number of cells in the G0/G1 stage of the cell cycle, but increased
granulosa cell viability. In summary, hCG induced Plk2 and Plk3 expression in the rat ovary. Prostaglandins and the
EGF signaling pathway are involved in regulating Plk2 expression. The transcription factor Sp1 is important for Plk2
transcriptional up-regulation. Our findings suggest that the increase in Plk2 and Plk3 expression contributes to the cell
cycle arrest of granulosa cells which is important for the luteinization of granulosa cells during the periovulatory period.
~2 Citings
386. 2,2',4,4'-Tetrabromodiphenyl ether (BDE-47) decreases progesterone synthesis through cAMP-PKA pathway
and P450scc downregulation in mouse Leydig tumor cells
By Han Xiumei; Tang Rong; Chen Xiaojiao; Xu Bo; Qin Yufeng; Wu Wei; Hu Yanhui; Xu Bin; Song Ling; Xia Yankai; et
al
From Toxicology (2012), 302(1), 44-50, Language: English, Database: MEDLINE
Polybrominated diphenyl ethers (PBDEs) are commonly used as flame retardants in textiles, plastics and electronics
and represent a group of persistent environmental contaminants. They have been found to accumulate in human and
marine mammals. Previous studies have shown that PBDEs have endocrine-disrupting properties and reproductive
toxicity. However, the mechanisms under the reproductive disruptions are still not well understood. In this study, we
explored the effects of 2,2',4,4'-tetrabromodiphenyl ether (BDE-47) on progesterone biosynthesis and possible
mechanisms in mouse Leydig tumor cells (mLTC-1). Our results showed that BDE-47 could reduce progesterone
production and decrease the intracellular cAMP level induced by hCG or forskolin. These suggested that BDE-47
decreasing progesterone production in mLTC-1 cells may be associated with the decline of intracellular cAMP level.
Moreover, our data also indicated that the site G protein in cAMP-PKA pathway may be involved in this process.
Furthermore, the addition of cAMP analog, 8-Br-cAMP, could not reverse the decrease of progesterone biosynthesis,
indicating that a post-cAMP site (or sites) might be involved into the BDE-47-decreased progesterone production. In
addition, we found BDE-47 reduced the activity of P450 side chain cleavage enzyme (P450scc), which was companied
with the decline of P450scc mRNA and protein level in mLTC-1 cells. Put all together, these results suggested that
progesterone synthesis decrease induced by BDE-47 may be associated with attenuation of cAMP generation and
reduction of P450scc activity.
~1 Citing
387. Linking αMSH with PPARγ in B16-F10 melanoma

By Maresca Vittoria; Flori Enrica; Camera Emanuela; Bellei Barbara; Aspite Nicaela; Ludovici Matteo; Catricala
Caterina; Cardinali Giorgia; Picardo Mauro
From Pigment cell & melanoma research (2013), 26(1), 113-27, Language: English, Database: MEDLINE
We have discovered a new α-melanocyte stimulating hormone (α-MSH)/peroxisome proliferator activated receptor-γ
(PPAR-γ) connection in B16-F10 cells. Both PPAR-γ up-regulation and its induction as an active transcription factor
were observed in response to α-MSH. The α-MSH/PPAR-γ connection influenced both pigmentation and proliferation.
The forskolin-stimulated cAMP/PKA pathway was not able to induce either PPAR-γ translocation into the nucleus or
PPAR-γ transcriptional activity. As the melanocortin-1 receptor, the specific receptor for the α-MSH, is a G-protein
coupled receptor, we wondered whether the phosphatidylinositol [PI(4,5)P(2) /PLC(β) ] signal pathway was involved in
mediating the α-MSH-dependent PPAR-γ activation. Employing inhibitors of PI(4,5)P(2) /PLC(β) pathway, the results
of our experiments suggested that this pathway was promoted by α-MSH and that α-MSH played a role in mediating
PPAR-γ activation. We have demonstrated, for the first time, that α-MSH induces the PI(4,5)P(2) /PLC(β) pathway,
through analysis of the basic steps of the pathway. The α-MSH effect on PPAR-γ was independent of animal species
and was not correlated with the physio-pathological status.
~2 Citings
388. Store-independent pathways for cytosolic STIM1 clustering in the regulation of store-operated Ca(2+) influx
By Zeng Bo; Chen Gui-Lan; Xu Shang-Zhong
STIM1 is a Ca(2+) sensing molecule. Once the Ca(2+) stores are depleted, STIM1 moves towards the plasma
membrane (PM) (translocation), forms puncta (clustering), and triggers store-operated Ca(2+) entry (SOCE). Although
this process has been regarded as a main mechanism for store-operated Ca(2+) channel activation, the STIM1
clustering is still unclear. Here we discovered a new phenomenon of STIM1 clustering, which is not triggered by
endoplasmic reticulum (ER) Ca(2+) depletion. STIM1 subplasmalemmal translocation and clustering can be induced
by ER Ca(2+) store depletion with thapsigargin (TG), G-protein-coupled receptor activator trypsin and ryanodine
receptor (RyR) agonists caffeine and 4-chloro-3-ethylphenol (4-CEP) in the HEK293 cells stably transfected with
STIM1-EYFP. The STIM1 clustering induced by TG was more sustained than that induced by trypsin and RyR
agonists. Interestingly, 4-CEP-induced STIM1 clustering also happened in the cytosol without ER Ca(2+) store
depletion. Application of some pharmacological regulators including flufenamic acid, 2-APB, and carbonyl cyanide 4-
(trifluoromethoxy)phenylhydrazone (FCCP) at concentrations without affecting ER Ca(2+) store also evoked cytosolic
STIM1 clustering. However, the direct store-operated ORAI channel blockers (SKF-96365, Gd(3+) and
diethylstilbestrol) or the signaling pathway inhibitors (genistein, wortmannin, Y-27632, forskolin and GF109203X) did
not change the STIM1 movement. Disruption of cytoskeleton by colchicine and cytochalasin D also showed no effect
on STIM1 movement. We concluded that STIM1 clustering and translocation are two dynamic processes that can be
pharmacologically dissociated. The ER Ca(2+) store-independent mechanism for STIM1 clustering is a new alternative
mechanism for regulating store-operated channel activity, which could act as a new pharmacological target.
~7 Citings
389. Canonical and noncanonical Hedgehog pathway in the pathogenesis of multiple myeloma
By Blotta Simona; Jakubikova Jana; Calimeri Teresa; Roccaro Aldo M; Amodio Nicola; Azab Abdel Kareem; Foresta
Umberto; Mitsiades Constantine S; Rossi Marco; Todoerti Katia; et al
From Blood (2012), 120(25), 5002-13, Language: English, Database: MEDLINE
The Hedgehog (Hh) pathway is required for cell-fate determination during the embryonic life, as well as cell growth and
differentiation in the adult organism, where the inappropriate activation has been implicated in several cancers. Here
we demonstrate that Hh signaling plays a significant role in growth and survival of multiple myeloma (MM) cells. We
observed that CD138(+) MM cells express Hh genes and confirmed Smoothened (Smo)-dependent Hh signaling in MM
using a novel synthetic Smo inhibitor, NVP-LDE225 (Novartis), which decreased MM cell viability by inducing specific
down-regulation of Gli1 and Ptch1, hallmarks of Hh activity. In addition, we detected a nuclear localization of Gli1 in
MM cells, which is completely abrogated by Forskolin, a Gli1-modulating compound, confirming Smo-independent
mechanisms leading to Hh activation in MM. Finally, we identified that bone marrow stromal cells are a source of the
Shh ligand, although they are resistant to the Hh inhibitor because of defective Smo expression and Ptch1 up-
regulation. Further in vitro as well as in vivo studies showed antitumor efficacy of NVP-LDE225 in combination with
bortezomib. Altogether, our data demonstrate activation of both canonical and noncanonical Hh pathway in MM, thus
providing the rationale for testing Hh inhibitors in clinical trials to improve MM patient outcome.
~19 Citings

390. Deficiency of proton-sensing ovarian cancer G protein-coupled receptor 1 attenuates glucose-stimulated
insulin secretion
By Nakakura Takashi; Mogi Chihiro; Tobo Masayuki; Tomura Hideaki; Sato Koichi; Kobayashi Masaki; Ohnishi Hiroshi;
Tanaka Shigeyasu; Wayama Mitsutoshi; Sugiyama Tetsuya; et al
Ovarian cancer G protein-coupled receptor 1 (OGR1) has been shown as a receptor for protons. In the present study,
we aimed to know whether OGR1 plays a role in insulin secretion and, if so, the manner in which it does. To this end,
we created OGR1-deficient mice and examined insulin secretion activity in vivo and in vitro. OGR1 deficiency reduced
insulin secretion induced by glucose administered ip, although it was not associated with glucose intolerance in vivo.
Increased insulin sensitivity and reduced plasma glucagon level may explain, in part, the unusual normal glucose
tolerance. In vitro islet experiments revealed that glucose-stimulated insulin secretion was dependent on extracellular
pH and sensitive to OGR1; insulin secretion at pH 7.4 to 7.0, but not 8.0, was significantly suppressed by OGR1
deficiency and inhibition of G(q/11) proteins. Insulin secretion induced by KCl and tolbutamide was also significantly
inhibited, whereas that induced by several insulin secretagogues, including vasopressin, a glucagon-like peptide 1
receptor agonist, and forskolin, was not suppressed by OGR1 deficiency. The inhibition of insulin secretion was
associated with the reduction of glucose-induced increase in intracellular Ca(2+) concentration. In conclusion, the
OGR1/G(q/11) protein pathway is activated by extracellular protons existing under the physiological extracellular pH of
7.4 and further stimulated by acidification, resulting in the enhancement of insulin secretion in response to high glucose
concentrations and KCl.
~4 Citings
391. Functional investigation of β-adrenoceptors in human isolated detrusor focusing on the novel selective β3-
adrenoceptor agonist KUC-7322
By Igawa Yasuhiko; Schneider Tim; Yamazaki Yoshinobu; Tatemichi Satoshi; Homma Yukio; Nishizawa Osamu;
Michel Martin C
From Naunyn-Schmiedeberg's archives of pharmacology (2012), 385(8), 759-67, Language: English, Database:
MEDLINE
This study aimed to characterize the β-adrenoceptor (β-AR) subtype mediating relaxation of isolated human bladder
strips and to explore relaxation by the novel β3-AR-selective agonist KUC-7322 for its relaxant effect on the human
isolated detrusor and for its effect on the carbachol (CCh)-induced contractile response. In two parallel studies,
relaxation of isolated human bladder strips was tested for the β-AR agonists isoproterenol, clenbuterol, BRL 37344,
and KUC-7322. For the isoproterenol and KUC-7322 responses, antagonism by CGP 20712A, ICI 118551, and
SR59230A was determined. The potency and efficacy of the reference agonists for detrusor relaxation was in line with
their known β3-AR activity. KUC-7322 relative to isoproterenol was a full agonist with a pEC(50) of 5.95 ± 0.09 and
5.92 ± 0.11 in the two studies. SR59230A exhibited antagonism of the expected potency against isoproterenol
(apparent pK (B) 7.2) but not against KUC-7322. Neither isoproterenol nor KUC-7322 nor forskolin significantly
attenuated CCh-induced contraction. These results suggest that KUC-7322 displays full agonistic activity in relaxing
the human detrusor without inhibiting the contraction induced by cholinergic stimulation. These characteristics, if
proven in vivo, may be beneficial for the treatment of overactive bladder, as increased bladder capacity with a
negligible effect on voiding contractions may be anticipated.
~8 Citings
392. Phosphorylation of cyclic AMP-response element-binding protein (CREB) is influenced by melatonin

treatment in pancreatic rat insulinoma β-cells (INS-1)
By Bazwinsky-Wutschke Ivonne; Wolgast Sabine; Muhlbauer Eckhard; Albrecht Elke; Peschke Elmar
From Journal of pineal research (2012), 53(4), 344-57, Language: English, Database: MEDLINE
The pineal hormone melatonin exerts its influence on the insulin secretion of pancreatic islets by a variety of signalling
pathways. The purpose of the present study was to analyse the impact of melatonin on the phosphorylated
transcription factor cAMP-response element-binding protein (pCREB). In pancreatic rat insulinoma β-cells (INS-1),
pCREB immunofluorescence intensities in cell nuclei using digitised confocal image analysis were measured to semi-
quantify differences in the pCREB immunoreactivity (pCREB-ir) caused by different treatments. Increasing
concentrations of forskolin or 3-isobutyl-1-methylxanthine (IBMX) resulted in a dose-dependent rise of the mean
fluorescence intensity in pCREB-ir nuclear staining. Concomitant melatonin application significantly decreased
pCREB-ir in INS-1 cells after 30-min, 1-hr and 3-hr treatment. The melatonin receptor antagonists luzindole and 4-
phenyl-2-propionamidotetraline (4P-PDOT) completely abolished the pCREB phosphorylation-decreasing effect of
melatonin, indicating that both melatonin receptor isoforms (MT(1) and MT(2)) are involved. In a transfected INS-1 cell
line expressing the human MT(2) receptor, melatonin caused the greatest reduction in pCREB after IBMX treatment
compared with nontransfected INS-1 cells, indicating a crucial influence of melatonin receptor density on pCREB
regulation. Furthermore, the downregulation of pCREB by melatonin is concomitantly associated with a statistically
significant downregulation of Camk2d transcript levels, as measured after 3 hr. In conclusion, the present study
provides evidence that the phosphorylation level of CREB is modulated in pancreatic β-cells by melatonin. Mediated
via CREB, melatonin regulates the expression of genes that play an important functional role in the regulation of β-cell
signalling pathways.
~6 Citings
393. The cAMP signaling system inhibits the repair of γ-ray-induced DNA damage by promoting Epac1-mediated
proteasomal degradation of XRCC1 protein in human lung cancer cells
By Cho Eun-Ah; Juhnn Yong-Sung
MEDLINE
Cyclic AMP is involved in the regulation of metabolism, gene expression, cellular growth and proliferation. Recently,
the cAMP signaling system was found to modulate DNA-damaging agent-induced apoptosis by regulating the
expression of Bcl-2 family proteins and inhibitors of apoptosis. Thus, we hypothesized that the cAMP signaling may
modulate DNA repair activity, and we investigated the effects of the cAMP signaling system on γ-ray-induced DNA
damage repair in lung cancer cells. Transient expression of a constitutively active mutant of stimulatory G protein
(GαsQL) or treatment with forskolin, an adenylyl cyclase activator, augmented radiation-induced DNA damage and
inhibited repair of the damage in H1299 lung cancer cells. Expression of GαsQL or treatment with forskolin or
isoproterenol inhibited the radiation-induced expression of the XRCC1 protein, and exogenous expression of XRCC1
abolished the DNA repair-inhibiting effect of forskolin. Forskolin treatment promoted the ubiquitin and proteasome-
dependent degradation of the XRCC1 protein, resulting in a significant decrease in the half-life of the protein after γ-ray
irradiation. The effect of forskolin on XRCC1 expression was not inhibited by PKA inhibitor, but 8-pCPT-2'-O-Me-
cAMP, an Epac-selective cAMP analog, increased ubiquitination of XRCC1 protein and decreased XRCC1 expression.
Knockdown of Epac1 abolished the effect of 8-pCPT-2'-O-Me-cAMP and restored XRCC1 protein level following γ-ray
irradiation. From these results, we conclude that the cAMP signaling system inhibits the repair of γ-ray-induced DNA
damage by promoting the ubiquitin-proteasome dependent degradation of XRCC1 in an Epac-dependent pathway in
lung cancer cells.
~4 Citings
394. Melanocortin 2 receptor-associated protein (MRAP) and MRAP2 in human adrenocortical tissues: regulation
of expression and association with ACTH responsiveness
By Hofland Johannes; Delhanty Patric J; Steenbergen Jacobie; Hofland Leo J; van Koetsveld Peter M; van Nederveen
Francien H; de Herder Wouter W; Feelders Richard A; de Jong Frank H
MEDLINE
CONTEXT: ACTH stimulates adrenocortical steroid production through the melanocortin 2 receptor (MC2R). MC2R
trafficking and signaling are dependent on the MC2R accessory protein (MRAP). The MRAP homolog MRAP2 also
transports the MC2R to the cell surface but might prevent activation. OBJECTIVE: The objective of the investigation
was to study the regulatory pathways of MRAP and MRAP2 and their contributions to ACTH responsiveness in human
adrenal tissues. DESIGN AND SETTING: MRAP, MRAP2, and MC2R expression levels were studied in 32 human
adrenocortical samples. Regulation of these mRNAs was investigated in 43 primary adrenal cultures, stimulated with
ACTH, forskolin, angiotensin II (AngII), phorbol-12-myristate-13-acetate (PMA), or dexamethasone. The induction of
cortisol, cAMP, and ACTH-responsive genes after treatment with ACTH was related to MRAP, MRAP2, and MC2R
expression levels. RESULTS: MRAP and MRAP2 levels were lower in adrenocortical carcinomas (ACC) than in other
adrenal tissues (P < 0.001). Patient ACTH and cortisol levels were associated with adrenal levels of MRAP and MC2R
in adrenal hyperplasia samples (P < 0.05) but not in tumors. ACTH induced the expression of MRAP 11 ± 2.1-fold and
MC2R 20 ± 3.8-fold in all adrenal tissue types (mean ± SEM, both P < 0.0001), whereas AngII augmented these
mRNAs 4.0 ± 1.2-fold and 12.6 ± 3.2-fold (P < 0.0001) in all but the ACC. MRAP2 expression was suppressed by
forskolin (-24 ± 15%, P = 0.013) and PMA (-22 ± 7%, P = 0.0007). MRAP, MRAP2, or MC2R levels were not
associated with the induction of cortisol, cAMP, or gene expression by ACTH in vitro. CONCLUSION: MRAP and
MC2R expression is induced by ACTH and AngII, which would facilitate cell surface receptor availability. Physiological
expression levels of MRAP, MRAP2, and MC2R were not limiting for ACTH sensitivity.
~7 Citings
395. Oncogenic K-ras expression is associated with derangement of the cAMP/PKA pathway and forskolin-
reversible alterations of mitochondrial dynamics and respiration
By Palorini R; De Rasmo D; Gaviraghi M; Sala Danna L; Signorile A; Cirulli C; Chiaradonna F; Alberghina L; Papa S
From Oncogene (2013), 32(3), 352-62, Language: English, Database: MEDLINE
The Warburg effect in cancer cells has been proposed to involve several mechanisms, including adaptation to hypoxia,
oncogenes activation or loss of oncosuppressors and impaired mitochondrial function. In previous papers, it has been
shown that K-ras transformed mouse cells are much more sensitive as compared with normal cells to glucose
withdrawal (undergoing apoptosis) and present a high glycolytic rate and a strong reduction of mitochondrial complex I.
Recent observations suggest that transformed cells have a derangement in the cyclic adenosine
monophosphate/cAMP-dependent protein kinase (cAMP/PKA) pathway, which is known to regulate several
mitochondrial functions. Herein, the derangement of the cAMP/PKA pathway and its impact on transformation-linked
changes of mitochondrial functions is investigated. Exogenous stimulation of PKA activity, achieved by forskolin
treatment, protected K-ras-transformed cells from apoptosis induced by glucose deprivation, enhanced complex I
activity, intracellular adenosine triphosphate (ATP) levels, mitochondrial fusion and decreased intracellular reactive
oxygen species (ROS) levels. Several of these effects were almost completely prevented by inhibiting the PKA
activity. Short-time treatment with compounds favoring mitochondrial fusion strongly decreased the cellular ROS levels
especially in transformed cells. These findings support the notion that glucose shortage-induced apoptosis, specific of
K-ras-transformed cells, is associated to a derangement of PKA signaling that leads to mitochondrial complex I
decrease, reduction of ATP formation, prevalence of mitochondrial fission over fusion, and thereby opening new
approaches for development of anticancer drugs.
~11 Citings
396. Potassium channel mutant KCNJ5 T158A expression in HAC-15 cells increases aldosterone synthesis
By Oki Kenji; Plonczynski Maria W; Luis Lam Milay; Gomez-Sanchez Elise P; Gomez-Sanchez Celso E
Primary aldosteronism is the most common cause of secondary hypertension, most frequently due to an aldosterone-
producing adenoma or idiopathic hyperaldosteronism. Somatic mutations of the potassium channel KCNJ5 in the
region of the selectivity filter have been found in a significant number of aldosterone-producing adenomas. There are
also familial forms of primary aldosteronism, one of which, familial hyperaldosteronism type 3 which to date has been
found in one family who presented with a severe abnormality in aldosterone and 18-oxocortisol production and
hypertrophy and hyperplasia of the transitional zone of the adrenal cortex. In familial hyperaldosteronism type 3, there
is a genomic mutation causing a T158A change of amino acids within the selectivity filter region of the KCNJ5 gene.
We are reporting our studies demonstrating that lentiviral-mediated expression of a gene carrying the T158A mutation
of the KCNJ5 in the HAC15 adrenal cortical carcinoma cell line causes a 5.3-fold increase in aldosterone secretion in
unstimulated HAC15-KCNJ5 cells and that forskolin-stimulated aldosterone secretion was greater than that of
angiotensin II. Expression of the mutated KCNJ5 gene decreases plasma membrane polarization, allowing sodium
and calcium influx into the cells. The calcium channel antagonist nifedipine and the calmodulin inhibitor W-7 variably
inhibited the effect. Overexpression of the mutated KCNJ5 channel resulted in a modest decrease in HAC15 cell
proliferation. These studies demonstrate that the T158A mutation of the KCNJ5 gene produces a marked stimulation
in aldosterone biosynthesis that is dependent on membrane depolarization and sodium and calcium influx into the
HAC15 adrenal cortical carcinoma cells.
~28 Citings
397. Phospholipase C-related but catalytically inactive protein (PRIP) modulates synaptosomal-associated
protein 25 (SNAP-25) phosphorylation and exocytosis
By Gao Jing; Takeuchi Hiroshi; Zhang Zhao; Fukuda Mitsunori; Hirata Masato
Exocytosis is one of the most fundamental cellular events. The basic mechanism of the final step, membrane fusion, is
mediated by the formation of the SNARE complex, which is modulated by the phosphorylation of proteins controlled by
the concerted actions of protein kinases and phosphatases. We have previously shown that a protein phosphatase-1
(PP1) anchoring protein, phospholipase C-related but catalytically inactive protein (PRIP), has an inhibitory role in
regulated exocytosis. The current study investigated the involvement of PRIP in the phospho-dependent modulation of
exocytosis. Dephosphorylation of synaptosome-associated protein of 25 kDa (SNAP-25) was mainly catalyzed by
PP1, and the process was modulated by wild-type PRIP but not by the mutant (F97A) lacking PP1 binding ability in in
vitro studies. We then examined the role of PRIP in phospho-dependent regulation of exocytosis in cell-based studies
using pheochromocytoma cell line PC12 cells, which secrete noradrenalin. Exogenous expression of PRIP
accelerated the dephosphorylation process of phosphorylated SNAP-25 after forskolin or phorbol ester treatment of the
cells. The phospho-states of SNAP-25 were correlated with noradrenalin secretion, which was enhanced by forskolin
or phorbol ester treatment and modulated by PRIP expression in PC12 cells. Both SNAP-25 and PP1 were co-
precipitated in anti-PRIP immunocomplex isolated from PC12 cells expressing PRIP. Collectively, together with our
previous observation regarding the roles of PRIP in PP1 regulation, these results suggest that PRIP is involved in the
regulation of the phospho-states of SNAP-25 by modulating the activity of PP1, thus regulating exocytosis.
~7 Citings
398. Red blood cell aggregation changes are depanded on its initial value: Effect of long-term drug treatment
From Clinical hemorheology and microcirculation (2012), , Language: English, Database: MEDLINE
~0 Citings
399. Hypoxia and MITF control metastatic behaviour in mouse and human melanoma cells
By Cheli Y; Giuliano S; Fenouille N; Allegra M; Hofman V; Hofman P; Bahadoran P; Lacour J-P; Tartare-Deckert S;
Bertolotto C; et al
From Oncogene (2012), 31(19), 2461-70, Language: English, Database: MEDLINE
Melanomas are very aggressive neoplasms with notorious resistance to therapeutics. It was recently proposed that
the remarkable phenotypic plasticity of melanoma cells allows for the rapid development of both resistance to
chemotherapeutic drugs and invasive properties. Indeed, the capacity of melanoma cells to form distant metastases is
the main cause of mortality in melanoma patients. Therefore, the identification of the mechanism controlling melanoma
phenotype is of paramount importance. In the present report, we show that deletion of microphthalmia-associated
transcription factor (MITF), the master gene in melanocyte differentiation, is sufficient to increase the metastatic
potential of mouse and human melanoma cells. MITF silencing also increases fibronectin and Snail, two mesenchymal
markers that might explain the increased invasiveness in vitro and in vivo. Furthermore, ablation of this population by
Forskolin-induced differentiation or MITF-forced expression significantly decreases tumour and metastasis formation,
suggesting that eradication of low-MITF cells might improve melanoma treatment. Moreover, we demonstrate that a
hypoxic microenvironment decreases MITF expression through an indirect, hypoxia-inducible factor 1 (HIF1)α-
dependant transcriptional mechanism, and increases the tumourigenic and metastatic properties of melanoma cells.
We identified Bhlhb2, a new factor in melanoma biology, as the mediator of hypoxia/HIF1α inhibitory effect on MITF
expression. Our results reveal a hypoxia-HIF1α-BHLHB2-MITF cascade controlling the phenotypic plasticity in
melanoma cells and favouring metastasis development. Targeting this pathway might be helpful in the design of new
anti-melanoma therapies.
~37 Citings
400. Leukemia inhibitory factor regulates differentiation of trophoblastlike BeWo cells through the activation of
JAK/STAT and MAPK3/1 MAP kinase-signaling pathways
By Leduc Katy; Bourassa Vincent; Asselin Eric; Leclerc Pierre; Lafond Julie; Reyes-Moreno Carlos
It is well established that syncytium formation involves the fusion of mononucleated trophoblasts into a multinucleated
structure and the secretion of hormonal factors, such as human chorionic gonadotropin (hCG). These morphological
and biochemical changes are regulated by a plethora of ligands, which upon binding to specific receptors trigger the
activation of many signaling pathways, such as janus kinase/signal transducer and activator of transcription
(JAK/STAT) and the mitogen-activated protein (MAP) kinase extracellular signal-regulated kinases 1 and 2 (MAPK3/1).
We used the forskolin-induced syncytialization of trophoblastlike BeWo cells to characterize at the cellular level the
effect mediated by leukemia inhibitory factor (LIF) on trophoblast differentiation and to describe its action at the
molecular level. Forskolin induces both hCG secretion and BeWo cell syncytial fusion. Although LIF had no effect on
the undifferentiated state of the cells, the cytokine generated a strong reduction in forskolin-induced hCG release. In
contrast to its effect on hCG secretion, LIF exerts a synergistic effect toward forskolin-induced fusion. LIF reduced
hormonal production through a STAT1- and STAT3-dependent mechanism, whereas MAPK3/1 was not involved in this
process. However, both types of signaling molecules were required to mediate the action of LIF in forskolin-induced
cell fusion. These data provide novel insights into the regulation of trophoblast cell differentiation by LIF and describe
for the first time the molecular mechanism underlying the effect of the cytokine.
~5 Citings
401. Role of Epac and protein kinase A in thyrotropin-induced gene expression in primary thyrocytes
By van Staveren Wilma C G; Beeckman Sandrine; Tomas Gil; Dom Genevieve; Hebrant Aline; Delys Laurent; Vliem
Marjolein J; Tresallet Christophe; Andry Guy; Franc Brigitte; et al
From Experimental cell research (2012), 318(5), 444-52, Language: English, Database: MEDLINE
cAMP pathway activation by thyrotropin (TSH) induces differentiation and gene expression in thyrocytes. We
investigated which partners of the cAMP cascade regulate gene expression modulations: protein kinase A and/or the
exchange proteins directly activated by cAMP (Epac). Human primary cultured thyrocytes were analysed by
microarrays after treatment with the adenylate cyclase activator forskolin, the protein kinase A (PKA) activator 6-MB-
cAMP and the Epac-selective cAMP analog 8-pCPT-2'-O-Me-cAMP (007) alone or combined with 6-MB-cAMP.
Profiles were compared to those of TSH. Cultures treated with the adenylate cyclase- or the PKA activator alone or
the latter combined with 007 had profiles similar to those induced by TSH. mRNA profiles of 007-treated cultures were
highly distinct from TSH-treated cells, suggesting that TSH-modulated gene expressions are mainly modulated by
cAMP and PKA and not through Epac in cultured human thyroid cells. To investigate whether the Epac-Rap-RapGAP
pathway could play a potential role in thyroid tumorigenesis, the mRNA expressions of its constituent proteins were
investigated in two malignant thyroid tumor types. Modulations of this pathway suggest an increased Rap pathway
activity in these cancers independent from cAMP activation.
~2 Citings
402. Phosphodiesterase inhibitors control A172 human glioblastoma cell death through cAMP-mediated
activation of protein kinase A and Epac1/Rap1 pathways
By Moon Eun-Yi; Lee Geun-Hee; Lee Myung-Shik; Kim Hwan-Mook; Lee Jae-Wook
From Life sciences (2012), 90(9-10), 373-80, Language: English, Database: MEDLINE
AIMS: We investigated whether cAMP-mediated protein kinase A(PKA) and Epac1/Rap1 pathways differentially affect
brain tumor cell death using 4-(3-cyclopentyloxy-4-methoxyphenyl)-2-pyrrolidone(rolipram), specific phosphodiesterase
type IV(PDE IV) inhibitor. MAIN METHODS: A172 and U87MG human glioblastoma cells were used. Percentage of
cell survival was determined by MTT assay. PKA and Epac1/Rap1 activation was determined by western blotting and
pull-down assay, respectively. Cell cycle and hypodiploid cell formation were assessed by flow cytometry analysis.
KEY FINDINGS: Non-specific PDE inhibitors, isobutylmethylxanthine(IBMX) and theophylline reduce survival
percentage of A172 and U87MG cells. The expression of PDE4A and PDE4B was detected in A172 and U87MG cells.
Rolipram-treated A172 or U87MG cell survival was lower in the presence of forskolin, adenylate cyclase activator, than
that in its absence. Co-treatment with rolipram and forskolin also enhanced CREB phosphorylation on serine 133 that
was inhibited by H-89, PKA inhibitor and cAMP-responsive guanine nucleotide exchange factor 1(Epac1), a Rap GDP
exchange factor-mediated Rap1 activity in A172 cells. When A172 cells were treated with cell-permeable dibutyryl-
cAMP(dbcAMP), PKA activator or 8-(4-chloro-phenylthio)-2'-O-methyladenosine-3',5'-cyclic monophosphate(CPT),
Epac1 activator, basal level of cell death was increased and cell cycle was arrested at the phase of G2/M. Rolipram-
induced A172 cell death was also increased by the co-treatment with dbcAMP or CPT, but it was inhibited by the pre-
treatment with H-89. SIGNIFICANCE: These findings demonstrate that PKA and Epac1/Rap1 pathways could
cooperatively play a role in rolipram-induced brain tumor cell death. It suggests that rolipram might regulate
glioblastoma cell density through dual pathways of PKA- and Epac1/Rap1-mediated cell death and cell cycle arrest.
~4 Citings
403. MicroRNA 335 is required for differentiation of malignant glioma cells induced by activation of
cAMP/protein kinase A pathway
By Shu Minfeng; Zhou Yuehan; Zhu Wenbo; Zhang Haipeng; Wu Sihan; Chen Jingkao; Yan Guangmei
From Molecular pharmacology (2012), 81(3), 292-8, Language: English, Database: MEDLINE
Glioma is the most common malignant cancer affecting the central nerve system, with dismal prognosis.
Differentiation-inducing therapy is a novel strategy that has been preliminarily proved effective against malignant
glioma. We have reported previously that activation of cAMP/protein kinase A (PKA) pathway is capable of inducing
glioma cell differentiation, characterized by astrocyte-like shape and dramatic induction of astrocyte biomarker glial
fibrillary acidic protein (GFAP). However, little progress has been made on molecular mechanisms related. Here we
demonstrate that microRNA 335 (miR-335) is responsible for the glioma cell differentiation stimulated by activation of
cAMP/PKA pathway. In the cAMP elevator cholera toxin-induced differentiation model of rat C6 glioma cells, miR-335
was significantly up-regulated, which was mimicked by other typical cAMP/PKA pathway activators (e.g., forskolin,
dibutyryl-cAMP) and abolished by PKA-specific inhibitor (9R,10S,12S)-2,3,9,10,11,12-hexahydro-10-hydroxy-9-methyl-
1-oxo-9,12-epoxy-1H-diindolo[1,2,3-fg:3',2',1'-kl]pyrrolo[3,4-i] [1,6]benzodiazocine-10-carboxylic acid, hexyl ester
(KT5720). In an assay measuring gain and loss of miR-335 function, exogenetic miR-335 resulted in induction of
GFAP, whereas miR-335 specific inhibitor antagomir-335 violently blocked cholera toxin-induced GFAP up-regulation.
It is noteworthy that in human U87-MG glioma cells and human primary culture glioma cells, miR-335 also mediated
cholera toxin-induced differentiation. Taken together, our findings suggest that miR-335 is potently required for
differentiation of malignant glioma cells induced by cAMP/PKA pathway activation, and a single microRNA may act as
an important fate determinant to control the differentiation status of malignant gliomas, which has provided a new
insight into differentiation-inducing therapy against malignant gliomas.
~8 Citings
404. Methylseleninic acid is a novel suppressor of aromatase expression

By Gao Ruijuan; Zhao Lijuan; Liu Xichun; Rowan Brian G; Wabitsch Martin; Edwards Dean P; Nishi Yoshihiro; Yanase
Toshihiko; Yu Qun; Dong Yan
From The Journal of endocrinology (2012), 212(2), 199-205, Language: English, Database: MEDLINE
Elevated circulating estrogen levels, as a result of increased peripheral aromatization of androgens by aromatase,
have been indicated to underlie the association between obesity and a higher risk of breast cancer in postmenopausal
women. Although aromatase inhibitors have been used as a first-line therapy for estrogen receptor-positive breast
cancer in postmenopausal women, their potential as breast cancer chemopreventive agents has been limited due to
toxicities and high costs. It is therefore imperative to develop new aromatase-inhibiting/suppressing agents with lower
toxicities and lower costs for breast cancer chemoprevention, especially in obese postmenopausal women. The
expression of the aromatase gene, CYP19, is controlled in a tissue-specific manner by the alternate use of different
promoters. In obese postmenopausal women, increased peripheral aromatase is primarily attributed to the activity of
the glucocorticoid-stimulated promoter, PI.4, and the cAMP-stimulated promoter, PII. In the present study, we show
that methylseleninic acid (MSA), a second-generation selenium compound, can effectively suppress aromatase
activation by dexamethasone, a synthetic glucocorticoid, and forskolin, a specific activator of adenylate cyclase. Unlike
the action of aromatase inhibitors, MSA suppression of aromatase activation is not mediated via direct inhibition of
aromatase enzymatic activity. Rather, it is attributable to a marked downregulation of promoters PI.4- and PII-specific
aromatase mRNA expression, and thereby a reduction of aromatase protein. Considering the low-cost and low-toxicity
nature of MSA, our findings provide a strong rationale for the further development of MSA as a breast cancer
chemopreventive agent for obese postmenopausal women.
~2 Citings
405. Hydroxylated estrogens (2-OH-E2 AND 4-OH-E2) do not activate cAMP/PKA and ERK1/2 pathways activation
in a breast cancer MCF-7 cell line
By Kwiecinska P; Ptak A; Wrobel A; Gregoraszczuk E L
From Endocrine regulations (2012), 46(1), 3-12, Language: English, Database: MEDLINE
OBJECTIVE: The current study was undertaken to determine the involvement of cAMP/PKA and MAPK-mediated
signalling pathways in the regulation of cell proliferation by hydroxylated metabolites of 17β-estradiol (E2). METHODS:
MCF-7, human breast cancer cells, were cultured in phenol red-free DMEM and treated with 1 nM 2-OH-E2 or 4-OH-
E2. E2 was used as a positive control. Cell proliferation was measured using the BrdU incorporation assay. Cellular
levels of cAMP and PKA were determined using ELISA kits. ERK1/2 protein expression was evaluated by Western
Blot analysis. To determine the involvement of different intracellular pathways in the regulation of cell proliferation
appropriate activators or inhibitors were used. RESULTS: Hydroxylated estrogens, as E2, exhibited no influence on
cAMP accumulation and PKA activation. In concomitant treatments with forskolin, cell proliferation decreased to the
amount noted under the influence of forskolin alone. A PKA inhibitor (PKI) had no statistically significant effect on
proliferation stimulated by E2 and its hydroxylated metabolites. Phospho-ERK1/2 protein expression in cells stimulated
with E2, 2-OH-E2 and 4-OH-E2 was not significantly different from the control. However, co-treatment with both
PD98059 and E2 or its hydroxylated metabolites reversed the effect of tested compounds on cell proliferation.
CONCLUSIONS: We have shown that E2 hydroxylated metabolites do not activate cAMP/PKA in breast cancer cells
and confirm previously published data, which showed a lack of ERK1/2 activation in a breast cancer cell line. The
observed reversible action of PD98059 on cell proliferation can be explained by the fact that hydroxylated estrogens,
as E2, stimulate secretion of a number of growth factors, which affect MAPK activity, suggested by Lobenhofer et al.
(2000).
~0 Citings
406. The influence of Aspalathus linearis (Rooibos) and dihydrochalcones on adrenal steroidogenesis:
quantification of steroid intermediates and end products in H295R cells
By Schloms Lindie; Storbeck Karl-Heinz; Swart Pieter; Gelderblom Wentzel C A; Swart Amanda C
From The Journal of steroid biochemistry and molecular biology (2012), 128(3-5), 128-38, Language: English,
Database: MEDLINE
The steroid hormone output of the adrenal gland is crucial in the maintenance of hormonal homeostasis, with hormonal
imbalances being associated with numerous clinical conditions which include, amongst others, hypertension, metabolic
syndrome, cardiovascular disease, insulin resistance and type 2 diabetes. Aspalathus linearis (Rooibos), which has
been reported to aid stress-related symptoms linked to metabolic diseases, contains a wide spectrum of bioactive
phenolic compounds of which aspalathin is unique. In this study the inhibitory effects of Rooibos and the
dihydrochalcones, aspalathin and nothofagin, were investigated on adrenal steroidogenesis. The activities of both
cytochrome P450 17α-hydroxylase/17,20 lyase and cytochrome P450 21-hydroxylase were significantly inhibited in
COS-1 cells. In order to study the effect of these compounds in H295R cells, a human adrenal carcinoma cell line, a
novel UPLC-MS/MS method was developed for the detection and quantification of twenty-one steroid metabolites
using a single chromatographic separation. Under both basal and forskolin-stimulated conditions, the total amount of
steroids produced in H295R cells significantly decreased in the presence of Rooibos, aspalathin and nothofagin.
Under stimulated conditions, Rooibos decreased the total steroid output 4-fold and resulted in a significant reduction of
aldosterone and cortisol precursors. Dehydroepiandrosterone-sulfate levels were unchanged, while the levels of
androstenedione (A4) and 11β-hydroxyandrostenedione (11βOH-A4) were inhibited 5.5 and 2.3-fold, respectively.
Quantification of 11βOH-A4 showed this metabolite to be a major product of steroidogenesis in H295R cells and we
confirm, for the first time, that this steroid metabolite is the product of the hydroxylation of A4 by human cytochrome
P450 11β-hydroxylase. Taken together our results demonstrate that Rooibos, aspalathin and nothofagin influence
steroid hormone biosynthesis and the flux through the mineralocorticoid, glucocorticoid and androgen pathways, thus
possibly contributing to the alleviation of negative effects arising from elevated glucocorticoid levels.
~3 Citings
407. CFTR and TMEM16A are separate but functionally related Cl- channels
By Ousingsawat Jiraporn; Kongsuphol Patthara; Schreiber Rainer; Kunzelmann Karl
From Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and
pharmacology (2011), 28(4), 715-24, Language: English, Database: MEDLINE
Previous reports point out to a functional relationship of the cystic fibrosis transmembrane conductance regulator
(CFTR) and Ca(2+) activated Cl(-) channels (CaCC). Recent findings showing that TMEM16A forms the essential part
of CaCC, prompted us to examine whether CFTR controls TMEM16A. Inhibition of endogenous CaCC by activation of
endogenous CFTR was found in 16HBE human airway epithelial cells, which also express TMEM16A. In contrast,
CFBE airway epithelial cells lack of CFTR expression, but express TMEM16A along with other TMEM16-proteins.
These cells produce CaCC that is inhibited by overexpression and activation of CFTR. In HEK293 cells coexpressing
TMEM16A and CFTR, whole cell currents activated by IMBX and forskolin were significantly reduced when compared
with cells expressing CFTR only, while the halide permeability sequence of CFTR was not changed. Expression of
TMEM16A, but not of TMEM16F, H or J, produced robust CaCC, which that were inhibited by CaCCinh-A01 and
niflumic acid, but not by CFTRinh-172. TMEM16A-currents were attenuated by additional expression of CFTR, and
were completely abrogated when additionally expressed CFTR was activated by IBMX and forskolin. On the other
hand, CFTR-currents were attenuated by additional expression of TMEM16A. CFTR and TMEM16A were both
membrane localized and could be coimmunoprecipitated. Intracellular Ca(2+) signals elicited by receptor-stimulation
was not changed during activation of CFTR, while ionophore-induced rise in [Ca(2+)](i) was attenuated after
stimulation of CFTR. The data indicate that both CFTR and TMEM16 proteins are separate molecular entities that
show functional and molecular interaction.
~13 Citings
408. Activation of LTRs from different human endogenous retrovirus (HERV) families by the HTLV-1 tax protein
and T-cell activators
By Toufaily Chirine; Landry Sebastien; Leib-Mosch Christine; Rassart Eric; Barbeau Benoit
From Viruses (2011), 3(11), 2146-59, Language: English, Database: MEDLINE
Human endogenous retroviruses (HERVs) represent approximately 8% of our genome. HERVs influence cellular gene
expression and contribute to normal physiological processes such as cellular differentiation and morphogenesis.
HERVs have also been associated with certain pathological conditions, including cancer and neurodegenerative
diseases. As HTLV-1 causes adult T-cell leukemia and HTLV-1-associated myelopathy/tropical spastic paraparesis
(HAM/TSP) and has been shown to modulate host gene expression mainly through the expression of the powerful Tax
transactivator, herein we were interested in looking at the potential modulation capacity of HTLV-1 Tax on HERV
expression. In order to evaluate the promoter activity of different HERV LTRs, pHERV-LTR-luc constructs were co-
transfected in Jurkat T-cells with a Tax expression vector. Tax expression potently increased the LTR activity of
HERV-W8 and HERV-H (MC16). In parallel, Jurkat cells were also stimulated with different T-cell-activating agents
and HERV LTRs were observed to respond to different combination of Forskolin, bpV[pic] a protein tyrosine
phosphatase inhibitor, and PMA. Transfection of expression vectors for different Tax mutants in Jurkat cells showed
that several transcription factors including CREB appeared to be important for HERV-W8 LTR activation. Deletion
mutants were derived from the HERV-W8 LTR and the region from -137 to -123 was found to be important for LTR
response following Tax expression in Jurkat cells, while a different region was shown to be required in cells treated
with activators. Our results thus demonstrated that HTLV-1 Tax activates several HERV LTRs. This raises the
possibility that upregulated HERV expression could be involved in diseases associated with HTLV-1 infection.
~10 Citings
409. β-Adrenergic receptors (β-AR) regulate VEGF and IL-6 production by divergent pathways in high β-AR-
expressing breast cancer cell lines
By Madden Kelley S; Szpunar Mercedes J; Brown Edward B
Activation of β-adrenergic receptors (β-AR) drives proangiogenic factor production in several types of cancers. To
examine β-AR regulation of breast cancer pathogenesis, β-AR density, signaling capacity, and functional responses to
β-AR stimulation were studied in four human breast adenocarcinoma cell lines. β-AR density ranged from very low in
MCF7 and MB-361 to very high in MB-231 and in a brain-seeking variant of MB-231, MB-231BR. Consistent with β-AR
density, β-AR activation elevated cAMP in MCF7 and MB-361 much less than in MB-231 and MB-231BR.
Functionally, β-AR stimulation did not markedly alter vascular endothelial growth factor (VEGF) production by MCF7 or
MB-361. In the two high β-AR-expressing cell lines MB-231 and MB-231BR, β-AR-induced cAMP and VEGF
production differed considerably, despite similar β-AR density. The β(2)-AR-selective agonist terbutaline and the
endogenous neurotransmitter norepinephrine decreased VEGF production by MB-231, but increased VEGF production
by MB-231BR. Moreover, β(2)-AR activation increased IL-6 production by both MB-231 and MB-231BR. These
functional alterations were driven by elevated cAMP, as direct activation of adenylate cyclase by forskolin elicited
similar alterations in VEGF and IL-6 production. The protein kinase A antagonist KT5720 prevented β-AR-induced
alterations in MB-231 and MB-231BR VEGF production, but not IL-6 production. Conclusions β-AR expression and
signaling is heterogeneous in human breast cancer cell lines. In cells with high β-AR density, β-AR stimulation
regulates VEGF production through the classical β-AR-cAMP-PKA pathway, but this pathway can elicit directionally
opposite outcomes. Furthermore, in the same cells, β-AR activate a cAMP-dependent, PKA-independent pathway to
increase IL-6 production. The complexity of breast cancer cell β-AR expression and functional responses must be
taken into account when considering β-AR as a therapeutic target for breast cancer treatment.
~18 Citings
410. Gefitinib induces apoptosis in human glioma cells by targeting Bad phosphorylation
By Chang Cheng-Yi; Shen Chiung-Chyi; Su Hong-Lin; Chen Chun-Jung
From Journal of neuro-oncology (2011), 105(3), 507-22, Language: English, Database: MEDLINE
Gefitinib, a selective epidermal growth factor receptor tyrosine kinase inhibitor, is under clinical testing and use in
cancer patients, including glioma. However, the molecular mechanisms involved in gefitinib-mediated anticancer
effects against glioma remain largely uncharacterized. Gefitinib inhibits cell growth and induces apoptosis in human
glioma cells. Gefitinib also induces death of H4 cells with characteristics of the intrinsic apoptotic pathway, including
Bax mitochondrial translocation, mitochondrial outer membrane permeabilization, cytochrome c cytosolic release, and
caspase-9/caspase-3 activation. The importance of Bax in mediating gefitinib-induced apoptosis was confirmed by the
attenuation of apoptosis by Bax siRNA and Bax channel blocker. Gefitinib caused Bad dephosphorylation, particularly
in serine-112, and increased its binding preference to Bcl-2 and Bcl-xL. The dephosphorylation of Bad in gefitinib-
treated cells was accompanied by reduced intracellular cyclic AMP content and protein kinase A (PKA) activity.
Adenylyl cyclase activator forskolin attenuated, but PKA inhibitor H89 augmented, gefitinib-induced Bad
dephosphorylation, Bax mitochondrial translocation, caspase-9/caspase-3 activation, and viability loss. Intriguingly, a
nonselective protein phosphatase inhibitor okadaic acid alleviated gefitinib-induced alterations, except Bad
dephosphorylation. In parallel with the higher basal PKA activity, response of U87 cells to gefitinib treatment was
delayed and relatively resistant compared with that of H4 and T98G cells. Inactivation of PKA sensitized H4, T98G,
and U87 cells to gefitinib cytotoxicity, Bad dephosphorylation in serine-112, and caspase-9/caspase-3 activation. Our
findings suggest the involvement of the Bad/Bax signaling pathway in gefitinib-induced glioma apoptosis. Furthermore,
the inactivation of PKA was shown to play a role in triggering the proapoptotic function of Bad.
~6 Citings
411. Transepithelial ion transport is suppressed in hypoxic sinonasal epithelium

By Blount Angela; Zhang Shaoyan; Chestnut Michael; Hixon Brian; Skinner Daniel; Sorscher Eric J; Woodworth
Bradford A
From The Laryngoscope (2011), 121(9), 1929-34, Language: English, Database: MEDLINE
OBJECTIVES/HYPOTHESIS: Sinonasal respiratory epithelial mucociliary clearance is dependent on the
transepithelial transport of ions such as Cl(-) . The objectives of the present study were to investigate the role of
oxygen restriction in 1) Cl(-) transport across primary sinonasal epithelial monolayers, 2) expression of the apical Cl(-)
channels cystic fibrosis transmembrane conductance regulator (CFTR) and transmembrane protein 16A (TMEM16A),
and 3) the pathogenesis of chronic rhinosinusitis. STUDY DESIGN: In vitro investigation. METHODS: Murine nasal
septal epithelial (MNSE), wild type, and human sinonasal epithelial (HSNE) cultures were incubated under hypoxic
conditions (1% O(2) , 5% CO(2) ). Cultures were mounted in Ussing chambers for ion transport measurements. CFTR
and TMEM16A expression were measured using quantitative reverse-transcription polymerase chain reaction (RT-
PCR). RESULTS: The change in short-circuit current (∆I(SC) in microamperes per square centimeter) attributable to
CFTR (forskolin-stimulated) was significantly decreased due to a 12-hour hypoxia exposure in both MNSE (13.55 ±
0.46 vs. 19.23 ± 0.18) and HSNE (19.55 ± 0.56 vs. 25.49 ± 1.48 [control]; P < .05). TMEM16A (uridine triphosphate-
stimulated transport) was inhibited by 48 hours of hypoxic exposure in MNSE (15.92 ± 2.87 vs. 51.44 ± 3.71 [control];
P < .05) and by 12 hours of hypoxic exposure in HSNE (16.75 ± 0.68 vs. 24.15 ± 1.35 [control]). Quantitative RT-PCR
(reported as relative mRNA levels ± standard deviation) demonstrated significant reductions in both CFTR and
TMEM16A mRNA expression in MNSE and HSNE owing to airway epithelial hypoxia. CONCLUSIONS: Sinonasal
epithelial CFTR and TMEM16A-mediated Cl(-) transport and mRNA expression were robustly decreased in an
oxygen-restricted environment. These findings indicate that persistent hypoxia may lead to acquired defects in
sinonasal Cl(-) transport in a fashion likely to confer mucociliary dysfunction in chronic rhinosinusitis.
~13 Citings
412. Red blood cell aggregation changes are depended on its initial value: Effect of long-term drug treatment
From Clinical hemorheology and microcirculation (2011), 48(4), 231-40, Language: English, Database: MEDLINE
~0 Citings
413. Inactivation of membrane surface ecto-5'-nucleotidase by sodium nitroprusside in C6 glioma cells

By Kumasaka Tadanori; Matsuoka Isao; Mashiko Hirobumi; Niwa Shin-ichi; Kimura Junko
From Journal of pharmacological sciences (2011), 117(1), 45-53, Language: English, Database: MEDLINE
Ecto-5'-nucleotidase (NT5E), a predominant enzyme that produces extracellular adenosine from AMP, plays an
important role in a variety of physiological and pathophysiological processes. This study was performed to identify
agents that affect NT5E activity using C6 glioma cells. When cells were incubated with sodium nitroprusside (SNP),
phorbol 12-myristate 13-acetate, forskolin, lipopolysaccharide, or interferon-γ, only SNP inhibited NT5E activity in a
time- and concentration-dependent manner (IC(50) = 1.2 µM). The inhibitory effect of SNP was long-lasting even after
SNP washout; and its action was not mimicked by nitric oxide generating agents, 8-bromo cyclic GMP, ferricyanide,
ferrocyanide, or sodium cyanide. SNP did not change NT5E mRNA level or membrane surface protein expression.
Similar to SNP, Fe(2+) inhibited NT5E activity, but to a lesser extent. Although Fe(2+) is known to increase oxidative
stress, Fe(2+)-mediated oxidative stress was not involved in SNP inhibition of NT5E because the inhibition of NT5E by
SNP was not affected by superoxide dismutase and catalase. In contrast, addition of Zn(2+), an essential metal co-
factor of NT5E activity, prevented SNP from inhibiting NT5E. These results suggest that SNP disrupts a critical Zn(2+)-
dependent enzyme activity and might be useful as a pharmacological tool for inhibiting NT5E.
~0 Citings
414. Molecular mechanisms of A3 adenosine receptor-induced G1 cell cycle arrest and apoptosis in androgen-
dependent and independent prostate cancer cell lines: involvement of intrinsic pathway
By Aghaei Mahmoud; Panjehpour Mojtaba; Karami-Tehrani Fatemeh; Salami Siamak
From Journal of cancer research and clinical oncology (2011), 137(10), 1511-23, Language: English, Database:
MEDLINE
PURPOSE: A3 adenosine receptor has shown several physiological and pathological activities, including cell
proliferation and apoptosis in various cancer cell lines. This study is designed to investigate molecular mechanism and
apoptotic pathway of A3 adenosine receptor in DU-145, PC3 and LNcap-FGC10 human prostate cancer cells.
METHODS: The expression level of A3 adenosine receptor was examined using real-time RT-PCR. cAMP
concentration was also measured. MTT viability, cell counting and BrdU incorporation tests were used to study the cell
proliferation effect of IB-MECA. Cell cycle analysis, Annexin V-FITC staining, Hoechst 33258 staining, mitochondrial
membrane potential (∆ΨM), caspase-3 activity, Bcl-2 and Bax protein expression were used to detect apoptosis.
RESULT: A3 adenosine receptors mRNAs were detected at different levels. IB-MECA inhibited forskolin-stimulated
cAMP. IB-MECA at (1 µM) suppressed cell proliferation and induced G1 cell cycle arrest. Indeed, IB-MECA down-
regulated the expression of CDK4, cyclin D1 and up-regulated p53 expression. IB-MECA at (10-100 µM) induced
apoptosis. The activity of caspase-3 was also increased. Expression of Bcl-2 was decreased in response to IB-
MECA, while the expression of Bax protein was increased. The results showed a significant loss of ∆ΨM, in a dose-
dependent manner. CONCLUSION: This study introduces a possible mechanism through A3 adenosine receptor
activation. IB-MECA inhibited prostate cancer cells proliferation and induced G1 cell cycle arrest through p53,
Cdk4/cyclinD1 pathway. Apoptosis determined by characteristic morphological changes and increased in sub-G1
population. Loss of MMP, activation of caspase-3 and down-regulation of Bcl-2 expression indicated mitochondrial
signaling pathway that involved in the apoptosis.
~3 Citings
415. The flavonone naringenin inhibits chloride secretion in isolated colonic epithelia
By Collins Danielle; Kopic Sascha; Geibel John P; Hogan Aisling M; Medani Mekki; Baird Alan W; Winter Desmond C
From European journal of pharmacology (2011), 668(1-2), 271-7, Language: English, Database: MEDLINE
Studies investigating the activating and inhibitory actions of bioflavonoids on colonic function have yielded conflicting
results. At low concentrations, flavonoids may stimulate chloride secretion while at higher concentrations they may
have antisecretory actions in the colon. Naringenin (4',5,7-trihydroxyflavanone), found predominantly in citrus fruits,
confers a protective effect against colorectal cancer and is purported to modulate secretory function in colonic cell
lines. The aim of this study was to investigate the effects of naringenin on ion transport in rat and human colonic
mucosae. Naringenin inhibited basal and stimulated chloride secretion in rat and human colonic mucosae mounted in
Ussing chambers (IC(50) 330 µMol/L and 360 µMol/L respectively) and did not alter intracellular cAMP generation.
Naringenin inhibited chloride secretion in MQAE (N-(ethoxycarbonylmethyl)-6-methoxyquinolinium bromide) loaded
crypts stimulated with forskolin. In BCECF (2',7'-bis-(2-carboxyethyl)-5-(and 6)-carboxyfluorescein acetoxymethyl
ester) loaded crypts, naringenin caused an intracellular acidification (∆pH/min=0.05 ± 0.004) which was sensitive to the
Na-K-Cl co-transporter (NKCC) inhibitor bumetanide. In addition, the antisecretory effect of naringenin was not
inhibited by blockade of barium sensitive basolateral K(+) transporters or by inhibition of Na+/H(+) exchange by
amiloride. We propose that the antisecretory action of naringenin is due to inhibition of basolateral NKCC1 in rat and
human colon.
~4 Citings
416. Gene network inference and biochemical assessment delineates GPCR pathways and CREB targets in
small intestinal neuroendocrine neoplasia
By Drozdov Ignat; Svejda Bernhard; Gustafsson Bjorn I; Mane Shrikant; Pfragner Roswitha; Kidd Mark; Modlin Irvin M
Small intestinal (SI) neuroendocrine tumors (NET) are increasing in incidence, however little is known about their
biology. High throughput techniques such as inference of gene regulatory networks from microarray experiments can
objectively define signaling machinery in this disease. Genome-wide co-expression analysis was used to infer gene
relevance network in SI-NETs. The network was confirmed to be non-random, scale-free, and highly modular.
Functional analysis of gene co-expression modules revealed processes including 'Nervous system development',
'Immune response', and 'Cell-cycle'. Importantly, gene network topology and differential expression analysis identified
over-expression of the GPCR signaling regulators, the cAMP synthetase, ADCY2, and the protein kinase A,
PRKAR1A. Seven CREB response element (CRE) transcripts associated with proliferation and secretion: BEX1,
BICD1, CHGB, CPE, GABRB3, SCG2 and SCG3 as well as ADCY2 and PRKAR1A were measured in an independent
SI dataset (n = 10 NETs; n = 8 normal preparations). All were up-regulated (p<0.035) with the exception of SCG3
which was not differently expressed. Forskolin (a direct cAMP activator, 10(-5) M) significantly stimulated transcription
of pCREB and 3/7 CREB targets, isoproterenol (a selective ss-adrenergic receptor agonist and cAMP activator, 10(-5)
M) stimulated pCREB and 4/7 targets while BIM-53061 (a dopamine D(2) and Serotonin [5-HT(2)] receptor agonist,
10(-6) M) stimulated 100% of targets as well as pCREB; CRE transcription correlated with the levels of cAMP
accumulation and PKA activity; BIM-53061 stimulated the highest levels of cAMP and PKA (2.8-fold and 2.5-fold vs.
1.8-2-fold for isoproterenol and forskolin). Gene network inference and graph topology analysis in SI NETs suggests
that SI NETs express neural GPCRs that activate different CRE targets associated with proliferation and secretion. In
vitro studies, in a model NET cell system, confirmed that transcriptional effects are signaled through the
cAMP/PKA/pCREB signaling pathway and that a SI NET cell line was most sensitive to a D(2) and 5-HT(2) receptor
agonist BIM-53061.
~9 Citings
417. Cytotoxic T lymphocyte antigen-2 alpha induces apoptosis of murine T-lymphoma cells and cardiac
fibroblasts and is regulated by cAMP/PKA
By Zhang Lingzhi; Yun Hongruo; Murray Fiona; Lu Ruilin; Wang Lin; Hook Vivian; Insel Paul A
The mechanism of cAMP-promoted apoptosis is not well defined. In wild-type (WT) murine S49 lymphoma cells,
cAMP promotes apoptosis in a protein kinase A (PKA)-dependent manner. We find that treatment of WT S49 cells
with 8-CPT-cAMP prominently increases the expression (as determined by DNA microarray analysis, real-time PCR
and immunblotting) of cytotoxic T lymphocyte antigen-2α (CTLA-2α), a cathepsin L-like cysteine protease inhibitor. By
contrast, CTLA-2α expression is only slightly increased by 8-CPT-cAMP treatment of D-S49 cells, which lack
cAMP/PKA-promoted apoptosis. Raising endogenous cAMP (by use of forskolin or inhibition of phosphodiesterase
[PDE] 4) or a PKA-selective, but not an Epac-selective, cAMP analogue, increases CTLA-2α mRNA expression; PKA,
and not Epac, thus mediates the increase in CTLA-2α expression. An adenoviral CLTA-2α (Ad-CTLA-2α) construct
induces apoptosis and enhances cAMP-promoted apoptosis in WT S49 cells but such cells do not have an increase in
cathepsin L activity nor does a cathepsin L inhibitor alter cAMP-promoted apoptosis. 8-CPT-cAMP also increases
CTLA-2α expression and induces apoptosis in murine cardiac fibroblasts; knockdown of CTLA-2α expression by
siRNA blocks 8-CPT-cAMP-promoted apoptosis. Thus, cAMP increases CTLA-2α expression in murine lymphoma
and cardiac fibroblasts and this increase in CTLA-2α contributes to cAMP/PKA-promoted apoptosis by mechanisms
that are independent of the ability of CTLA-2α to inhibit cathepsin L.
~4 Citings
418. Berberine Reduces cAMP-Induced Chloride Secretion in T84 Human Colonic Carcinoma Cells through
Inhibition of Basolateral KCNQ1 Channels
By Alzamora Rodrigo; O'Mahony Fiona; Ko Wing-Hung; Yip Tiffany Wai-Nga; Carter Derek; Irnaten Mustapha; Harvey
Brian Joseph
From Frontiers in physiology (2011), 233, Language: English, Database: MEDLINE
Berberine is a plant alkaloid with multiple pharmacological actions, including antidiarrhoeal activity and has been
shown to inhibit Cl(-) secretion in distal colon. The aims of this study were to determine the molecular signaling
mechanisms of action of berberine on Cl(-) secretion and the ion transporter targets. Monolayers of T84 human
colonic carcinoma cells grown in permeable supports were placed in Ussing chambers and short-circuit current
measured in response to secretagogues and berberine. Whole-cell current recordings were performed in T84 cells
using the patch-clamp technique. Berberine decreased forskolin-induced short-circuit current in a concentration-
dependent manner (IC(50) 80 ± 8 µM). In apically permeabilized monolayers and whole-cell current recordings,
berberine inhibited a cAMP-dependent and chromanol 293B-sensitive basolateral membrane K(+) current by 88%,
suggesting inhibition of KCNQ1 K(+) channels. Berberine did not affect either apical Cl(-) conductance or basolateral
Na(+)-K(+)-ATPase activity. Berberine stimulated p38 MAPK, PKCα and PKA, but had no effect on p42/p44 MAPK
and PKCδ. However, berberine pre-treatment prevented stimulation of p42/p44 MAPK by epidermal growth factor.
The inhibitory effect of berberine on Cl(-) secretion was partially blocked by HBDDE (∼65%), an inhibitor of PKCα and
to a smaller extent by inhibition of p38 MAPK with SB202190 (∼15%). Berberine treatment induced an increase in
association between PKCα and PKA with KCNQ1 and produced phosphorylation of the channel. We conclude that
berberine exerts its inhibitory effect on colonic Cl(-) secretion through inhibition of basolateral KCNQ1 channels
responsible for K(+) recycling via a PKCα-dependent pathway.
~0 Citings
419. Beyond reverse pharmacology: Mechanism-based screening of Ayurvedic drugs

By Lele R D
From Journal of Ayurveda and integrative medicine (2010), 1(4), 257-65, Language: English, Database: MEDLINE
This paper reviews the pharmacology of Indian medicinal plants, starting with the historical background of European
work on the subject beginning as early as the 17th century, and tracing its history through the work of Sen and Bose in
the 1930's, and Vakhil's historic 1949 paper on Sarpaghanda. The often crucial role of patient feedback in early
discoveries is highlighted, as is the time lag between proof of pharmacological action and identification of the active
principle, and subsequent elucidation of mechanism of action. In the case of Indian plants in the 20th century this
process sometimes took almost 50 years. Reserpine and its mechanisms are given in detail, and its current relevance
to public health discussed. The foundation of present day methods of pharmacology is briefly presented so the
complexity of methods used to identify properties of Ayurveda derived drugs like forskolin and baicalein, and their
bioavailability, may be better appreciated. Ayurveda derived anti-oxidants and their levels of action, immuno-
modulators, particularly with respect to the NF-kB pathway and its implications for cancer control, are all considered.
The example of curcumin derived from turmeric is explained in more detail, because of its role in cancer prevention.
Finally, the paper emphasizes the importance of Ayurveda's concepts of rasayana as a form of dietary chemo-
prevention; the significance of ahar, diet, in Ayurveda's aspiration to prevent disease and restore health thus becomes
clear. Understood in this light, Ayurveda may transcend pharmacology as a treatment paradigm.
~1 Citing
420. Promoter-specific effects of metformin on aromatase transcript expression

By Samarajeewa Nirukshi U; Ham Seungmin; Yang Fangyuan; Simpson Evan R; Brown Kristy A
From Steroids (2011), 76(8), 768-71, Language: English, Database: MEDLINE
Phase III aromatase inhibitors (AIs) are proving successful in the treatment of hormone-dependent postmenopausal
breast cancer. Side-effects associated with total body aromatase inhibition have prompted new research into the
development of breast-specific AIs. The identification of tissue- and disease-specific usage of aromatase promoters
has made the inhibition of aromatase at the transcriptional level an interesting approach. We have previously
demonstrated that AMPK-activating drugs, including metformin, were potent inhibitors of aromatase expression in
primary human breast adipose stromal cells (hASCs). This study examines the promoter-specific effects of metformin
on inhibiting aromatase expression in hASCs. Tumour-associated promoters PII/PI.3 were activated using forskolin
(FSK)/phorbol ester (PMA), whereas normal adipose associated promoter PI.4 was activated using dexamethasone
(DEX)/tumour necrosis factor-α (TNFα). Results demonstrate that metformin significantly decreased the FSK/PMA-,
but not the DEX/TNFα-mediated expression of total aromatase at concentrations of 10, 20, and 50 µM (P ≤ 0.05).
Using PCR to amplify promoter-specific transcripts of aromatase, it appears that the inhibition of the FSK/PMA-
mediated expression of aromatase is due to decreases in PII/PI.3-specific transcripts, whereas no effect of metformin
is observed on any promoter-specific transcript, including PI.4, in DEX/TNFα-treated hASCs. This report therefore
supports the hypothesis that metformin would act as a breast-specific inhibitor of aromatase expression in the context
of postmenopausal breast cancer.
~5 Citings
421. Tissue-specific regulation of aromatase promoter II by the orphan nuclear receptor LRH-1 in breast adipose
stromal fibroblasts
By Chand Ashwini L; Herridge Kerrie A; Howard Tamara L; Simpson Evan R; Clyne Colin D
From Steroids (2011), 76(8), 741-4, Language: English, Database: MEDLINE
In postmenopausal breast cancers, the increase in aromatase expression observed in tumour associated adipose
stromal cells is mediated via the upregulation of promoter II (PII) transcription. Factors such as PGE2 which are
secreted from breast carcinomas induce PII expression. The orphan nuclear receptor LRH-1/NR5A2 is one of the
critical downstream transcriptional mediators of this effect. The aim of the current study was to determine whether
LRH-1 could bind directly to PII and whether the suppression of LRH-1 expression could inhibit aromatase expression
in human adipose stromal fibroblasts. Chromatin immunoprecipitation demonstrated endogenous LRH-1 occupancy
on PII under basal conditions and with treatment with forskolin and phorbol 12-myristate 13-acetate (PMA). To assess
the impact of LRH-1 knockdown on FSK/PMA mediated PII expression, cells were transfected with shRNA targeted
against LRH-1 (shLRH-1) and treated with forskolin and PMA. A decrease in LRH-1, PII and total aromatase mRNA
transcripts was observed in shLRH-1 transfected cells compared to controls under basal and treatment conditions.
The results of this study support the hypothesis that suppression of LRH-1 may potentially be beneficial in the tissue
specific regulation of aromatase expression in post menopausal breast cancer.
~5 Citings
422. Role of histamine H4 receptor in breast cancer cell proliferation

By Medina Vanina A; Brenzoni Pablo G; Lamas Diego J Martinel; Massari Noelia; Mondillo Carolina; Nunez Mariel A;
Pignataro Omar; Rivera Elena S
From Frontiers in bioscience (Elite edition) (2011), 31042-60, Language: English, Database: MEDLINE
In order to better understand the role of histamine H4 (H4R) receptor in breast cancer, we studied the receptor
expression pattern, associated signal transduction pathway and biological responses, in breast cancer cell lines with
different malignant characteristics. A different pattern of protein expression was observed in MDA-MB-231 compared
to MCF-7 cells determined by western blot, exhibiting the presence of a diverse range of molecular weight species of
the H4R. H4R agonist reduced cyclic adenosine monophosphate (cAMP) formation induced by forskolin only in MCF-7
cells. In MDA-MB-231 cells, H4R agonists significantly decreased cell proliferation, augmented the Annexin-V and
TdT-mediated UTP-biotin Nick End labelling (TUNEL) positive cells and produced a 2.5-fold increase in cell
senescence. In MCF-7 cells, H4R agonists inhibited proliferation by 50%, increasing the exponential doubling time.
This effect was associated to an augment in Annexin-V and TUNEL positive cells, and a 2-fold increase in cell
senescence. We conclude that H4R is functionally expressed in human breast cancer cell lines, exhibiting a key role in
histamine-mediated biological processes such as cell proliferation, senescence and apoptosis.
~7 Citings
423. Cyclic AMP-induced p53 destabilization is independent of EPAC in pre-B acute lymphoblastic leukemia
cells in vitro
By Safa Majid; Kazemi Ahmad; Zaker Farhad; Razmkhah Farnaz
From Journal of receptor and signal transduction research (2011), 31(3), 256-63, Language: English, Database:
MEDLINE
CONTEXT: Activation of the tumor suppressor protein p53 facilitates the cellular response to genotoxic stress. Thus,
releasing the wild-type p53 from indirect suppression would be crucial to successful killing of cancer cells by DNA-
damaging therapeutic agents. OBJECTIVE: The aim of this study was to investigate the inhibitory role of cyclic
adenosine monophosphate (cAMP) levels on p53 protein in acute lymphoblastic leukemia (ALL) cells. More
importantly, we were interested to show through which receptor cAMP acts to promote p53 degradation. MATERIALS
AND METHODS: In cell cultures, we investigated the effects of forskolin/3-isobutyl-1-methylxanthine (IBMX) on
stimulated p53 of ALL cell lines. Western blotting analysis was performed to detect the expression of p53, phospho-
p53, acetylated-p53, phospho-cAMP response element-binding protein (CREB), and Mdm2 proteins. Flow cytometry
was applied to analyze apoptosis. The gene expression of p53 and its target genes was examined by real-time
polymerase chain reaction. RESULTS: We show that elevation of cAMP levels in ALL cells exposed to DNA damage
attenuates p53 accumulation. Inhibition of proteosome function with MG-132 reversed the inhibitory effect of cAMP on
p53. However, targeting the p53-Mdm2 interaction did not rescue accumulated p53 from the destabilizing signal of
cAMP. The specific agonist of the cAMP receptor exchange protein activated by cAMP had no effect on p53
expression in doxorubicin-treated NALM-6 cells, whereas PKA activators decreased p53 accumulation. DISCUSSION
AND CONCLUSION: Our studies demonstrate that cAMP-PKA pathway regulates the sensitivity toward DNA-
damaging agents via inhibition of a p53-dependent pathway in B-cell precursor ALL (BCP-ALL) cells.
~4 Citings
424. Processing of proaugurin is required to suppress proliferation of tumor cell lines

By Ozawa Akihiko; Lick Adam N; Lindberg Iris
From Molecular endocrinology (Baltimore, Md.) (2011), 25(5), 776-84, Language: English, Database: MEDLINE
Augurin is a secretory molecule produced in pituitary, thyroid, and esophagus and implicated in a wide array of
physiological processes, from ACTH release to tumor suppression. However, the specific proaugurin-derived peptides
present in various cell types are not yet known. In order to shed light on the posttranslational modifications required for
biological activity, we here describe the posttranslational processing of proaugurin in AtT-20 and Lovo cells and identify
proaugurin-derived products generated by convertases. In vitro cleavage of proaugurin with proprotein convertases
produced multiple peptides, including a major product with a mass of 9.7 kDa by mass spectrometry. Metabolic
labeling of C-terminally tagged proaugurin in AtT-20 and AtT-20/PC2 cells resulted in a major 15-kDa tagged form on
SDS-PAGE, which likely corresponds to the 9.7-kDa in vitro fragment, with the added tag, its linker, and
posttranslational modification(s). The secretion of neither proaugurin nor this cleavage product was stimulated by
forskolin, indicating its lack of storage in regulated secretory granules and lack of cleavage by PC2. Incubation of cells
with the furin inhibitor nona-d-arginine resulted in impaired cleavage of proaugurin, whereas metalloprotease inhibitors
did not affect proaugurin proteolysis. These data support the idea that proaugurin is cleaved by furin and secreted via
the constitutive secretory pathway. Interestingly, proaugurin was sulfated during trafficking; sulfation was completely
inhibited by brefeldin A. Proliferation assays with three different tumor cell lines demonstrated that only furin-cleaved
proaugurin could suppress cell proliferation, suggesting that proteolytic cleavage is a posttranslational requirement for
proaugurin to suppress cell proliferation.
~15 Citings
425. Rapid differentiation of human embryonal carcinoma stem cells (NT2) into neurons for neurite outgrowth
analysis
By Tegenge Million Adane; Roloff Frank; Bicker Gerd
From Cellular and molecular neurobiology (2011), 31(4), 635-43, Language: English, Database: MEDLINE
Human neurons derived from stem cells can be employed as in vitro models to predict the potential of neurochemicals
affecting neurodevelopmental cellular processes including proliferation, migration, and differentiation. Here, we
developed a model of differentiating human neurons from well characterized human embryonal carcinoma stem cells
(NT2). NT2 cells were induced to differentiate into neuronal phenotypes after 2 weeks of treatment with retinoic acid in
aggregate culture. Nestin positive progenitor cells migrate out of NT2 aggregates and differentiate into βIII-tubulin
expressing neuronal cells. Culturing the NT2 cells for an additional 7-14 days resulted in increased percentage of βIII-
tubulin expressing cells, elaborating a long neurite that positively stained for axonal marker (Tau) and presynaptic
protein (synapsin). We then asked whether neurite outgrowth from NT2 cells is modulated by bioactive chemicals.
Since the cAMP/PKA pathway has been widely investigated as a regulator of neurite outgrowth/regeneration in several
experimental systems, we used chemical activators and inhibitors of cAMP/PKA pathway in the culture. The adenylyl
cyclase activator, forskolin, and cell-permeable analog of cAMP, 8-Br-cAMP increased the percentage of neurite
bearing cells and neurite extension. Application of the protein kinase A inhibitors, H-89 and Rp-cAMP, blocked neurite
formation. Taken together, NT2 aggregates undergo migration, differentiation, and neurite elaboration and can be
used as a model of differentiating human neurons to screen neurochemicals and to understand cellular mechanisms of
human nerve cell development.
~7 Citings
426. Expression of membrane and nuclear progesterone receptors in two human placental choriocarcinoma cell
lines (JEG-3 and BeWo): Effects of syncytialization
By Zachariades Elena; Foster Helen; Goumenou Anastasia; Thomas Peter; Rand-Weaver Mariann; Karteris
Emmanouil
From International journal of molecular medicine (2011), 27(6), 767-74, Language: English, Database: MEDLINE
A vital function of the human placenta is to produce steroid hormones such as progesterone, which are essential for
the maintenance of pregnancy and the onset of parturition. Although choriocarcinoma cell lines are valuable placental
models for investigations of steroid hormone actions, little is known about the expression of progesterone receptors
(PRs) in these cell lines. Therefore, in this study, the expression of membrane and nuclear PRs was investigated in
cultures of fusigenic (BeWo) and non-fusigenic (JEG-3) human choriocarcinoma cell lines. In addition, the effects of an
inducer of syncytialization (forskolin) on the PR expression in BeWo cells were assessed. Quantitative RT-PCR
revealed that in fully syncytialized BeWo cells (treated with 50 µM forskolin for 72 h) there was a significant down-
regulation of mPRα and up-regulation of mPRβ and of the progesterone membrane component-1 (PGRMC1) when
compared with non-syncytialized BeWo cells. Expression of all the mPR and PGRMC1 mRNAs was significantly lower
in JEG-3 cells compared to non-syncytialized BeWo cells. Interestingly, expression of PR-B was unaltered between
the two BeWo states but was significantly higher in JEG-3 cells. Immunofluorescence analysis revealed that mPR
proteins are differentially expressed in these choriocarcinoma cell lines as well as in the human placenta. The data
demonstrate that human choriocarcinoma cell lines have a complex system of progesterone signalling involving
multiple classes of PRs. The finding that syncytialization is accompanied by changes in the expression of these
receptors may suggest that this process influences progesterone signalling.
~1 Citing
427. Forskolin, a Hedgehog signal inhibitor, inhibits cell proliferation and induces apoptosis in pediatric tumor
cell lines
By Yamanaka Hiroaki; Oue Takaharu; Uehara Shuichiro; Fukuzawa Masahiro
From Molecular medicine reports (2010), 3(1), 133-9, Language: English, Database: MEDLINE
The Hedgehog (Hh) signaling pathway regulates the development of many organs in mammals. Recent studies have
indicated that the activation of the Hh signaling pathway contributes to the growth of various adult cancers. However,
little is known about its role in the development of pediatric malignancies. The present study was undertaken to
examine the expression and functional involvement of Hh signal transcription factors in pediatric tumor cells in order to
determine their potential as therapeutic targets. We utilized real-time RT-PCR to investigate the expression of Glioma-
associated oncogene homolog 1 (Gli1) in various pediatric tumor cell lines, including rhabdomyosarcoma,
neuroblastoma and hepatoblastoma. The mRNA expression of Gli1 was markedly increased in rhabdomyosarcoma
(RMS-YM, RD, RH30) cell lines, and moderately increased in neuroblastoma (NB19) and hepatoblastoma (Huh6) cell
lines. The proliferation of these cell lines was dose dependently inhibited by Forskolin, a specific Hh signal inhibitor. In
addition, Forskolin-induced growth suppression was associated with the down-regulation of C-Myc. Moreover, the
blockade of Hh signaling with Forskolin enhanced cell apoptosis in a dose-dependent manner. These results
demonstrated that Hh signal activation frequently occurs in neuroblastoma, hepatoblastoma and rhabdomyosarcoma
cell lines. The inhibition of Hh signaling suppressed proliferation and increased apoptosis in these tumor cells. These
findings suggest that the Hh signaling pathway plays an important role in tumorigenesis and is a potential molecular
target of new treatment strategies for these pediatric malignant tumors.
~0 Citings
428. Fibronectin increases RhoA activity through inhibition of PKA in the human gastric cancer cell line SGC-
7901
By Li Yongjin; Chen Yongchang; Tao Yan; Wang Ying; Chen Yuefang; Xu Wenrong
From Molecular medicine reports (2011), 4(1), 65-9, Language: English, Database: MEDLINE
Fibronectin/integrin-mediated signaling plays a key role in the regulation of adhesion, migration and metastasis of
tumors. Numerous studies have addressed the significance of the association between integrin and RhoA, but the
exact mechanism is unclear. Results from laboratories, including ours, have demonstrated that PKA inhibits the
activity and function of RhoA. This study was designed to investigate the relationships among the fibronectin/integrin-,
cAMP/PKA- and RhoA-mediated intracellular signal transduction pathways. Rho activity was detected by pull-down
assay. cAMP concentration was measured by radioimmunoassay. The distribution of the PKA catalytic subunit and the
phosphorylation of vasodilator-stimulated phosphoprotein (VASP) were detected by fluorescence microscopy and
Western blotting, respectively, to examine the activation of PKA. cAMP-mediated gene expression activity was
analyzed using a luciferase reporter gene assay. The results revealed that, in SGC-7901 cells, soluble fibronectin
increased RhoA activity and blocked the inhibition of RhoA activity by cAMP/PKA. The cAMP level, which was
increased by forskolin and pertussis toxin, was decreased by fibronectin. The nuclear location of the PKA catalytic
unit, the phosphorylation of VASP and cAMP response element (CRE)-directed reporter gene expression induced by
forskolin were blocked by fibronectin. However, fibronectin did not block VASP phosphorylation or CRE-directed
reporter gene expression induced by cAMP. These data suggest that fibronectin/integrin induces RhoA activation
through the inhibition of cAMP/PKA signal transduction. The possible point of action of fibronectin/integrin is adenylate
cyclase.
~1 Citing
429. Norepinephrine induces VEGF expression and angiogenesis by a hypoxia-inducible factor-1α protein-
dependent mechanism
By Park Soon Young; Kang Joo Hee; Jeong Kang Jin; Lee Jangsoon; Han Jeong Whan; Choi Wahn Soo; Kim Yong
Kee; Kang Jaeku; Park Chang Gyo; Lee Hoi Young
From International journal of cancer (2011), 128(10), 2306-16, Language: English, Database: MEDLINE
A growing number of studies have demonstrated that physiological factors can influence the progression of several
cancers via cellular immune function, angiogenesis and metastasis. Recently, stress-induced catecholamines have
been shown to increase the expression of various cancer progressive factors, including vascular endothelial growth
factor (VEGF), matrix metalloproteinases and interleukins. However, a detailed mechanism remains to be identified.
In this study, we investigated the role of adrenergic receptors and hypoxia-inducible factor (HIF)-1α protein in
catecholamine-induced VEGF expression and angiogenesis. Treatment of the cells with norepinephrine (NE) or
isoproterenol induced VEGF expression and HIF-1α protein amount in a dose-dependent manner. Induction of VEGF
expression by NE was abrogated when the cells were transfected with HIF-1α-specific siRNA. Similarly, adenylate
cyclase activator forskolin and cyclic AMP-dependent protein kinase A inhibitor H-89 enhanced and decreased HIF-1α
protein amount, respectively. More importantly, conditioned medium of NE-stimulated cancer cells induced
angiogenesis in a HIF-1α protein-dependent manner. In addition, pretreatment of cells with propranolol, a β-adrenergic
receptor (AR) blocker, completely abolished induction of VEGF expression and HIF-1α protein amount by NE in all of
the tested cancer cells. However, treatment with the α1-AR blocker prazosin inhibited NE-induced HIF-1α protein
amount and angiogenesis in SK-Hep1 and PC-3 but not MDA-MB-231 cells. Collectively, our results suggest that ARs
and HIF-1α protein have critical roles in NE-induced VEGF expression in cancer cells, leading to stimulation of
angiogenesis. These findings will help to understand the mechanism of cancer progression by stress-induced
catecholamines and design therapeutic strategies for cancer angiogenesis.
~21 Citings
430. Heterotrimeric stimulatory GTP-binding proteins inhibit cisplatin-induced apoptosis by increasing X-linked
inhibitor of apoptosis protein expression in cervical cancer cells
By Cho Eun-Ah; Oh Jung-Min; Kim So-Young; Kim Yeni; Juhnn Yong-Sung
From Cancer science (2011), 102(4), 837-44, Language: English, Database: MEDLINE
Treatment with cisplatin (cis-dichlorodiammineplatinum (II)) induces DNA double-stranded breaks and apoptosis in
many human cancer cells. We have reported that heterotrimeric stimulatory GTP-binding proteins (Gαs) can modulate
the apoptotic response of several cancer cells. This study investigated the effect of Gαs on apoptosis triggered by
cisplatin and its underlying molecular mechanism in cervical cancer cells. Stable expression of constitutively active
Gαs (GαsQL) decreased the release of cytochrome c from the mitochondria to the cytosol and cleavage of caspase-3
and poly(ADP-ribose) polymerases in HeLa cells treated with 30 µM cisplatin, indicating that Gαs inhibited cisplatin-
induced apoptosis. Treatment with forskolin also inhibited apoptosis of C33A and CaSKi cervical cancer cells.
Expression of GαsQL increased the expression of the X-linked inhibitor of apoptosis protein (XIAP) and partially
maintained increased XIAP after cisplatin treatment. Knockdown of XIAP by siRNA augmented apoptosis. Expression
of GαsQL increased XIAP mRNA; this increase was inhibited by a protein kinase A inhibitor and cAMP response
element (CRE) decoy. A cAMP response element (CRE)-like element at -1396 bp in the XIAP promoter was found to
mediate the induction of XIAP by Gαs. In addition, expression of GαsQL protected against the ubiquitin/proteasome-
dependent degradation of the XIAP protein. This study shows that Gαs inhibits cisplatin-induced apoptosis by
increasing transcription of XIAP and by decreasing degradation of XIAP protein in HeLa cervical cancer cells.
~4 Citings
431. Down-regulation of ICBP90 contributes to doxorubicin resistance

By Wang Jingxuan; Song Ying; Xu Shanqi; Zhang Qingyuan; Li Yulian; Tang Dabei; Jin Shi
Acquired resistance to doxorubicin has become a serious obstacle in breast cancer treatment. The underlying
mechanism responsible for this has not been completely elucidated. In this study, a doxorubicin-resistant MCF-7/Dox
cell was developed to mimic the occurrence of acquired doxorubicin resistance. We next contrasted the expression
profiles of ICBP90 and Topo IIα and tumor cell growth of different breast cancer cell lines to doxorubicin. Decreased
expression levels of ICBP90 and Topo IIα were found in doxorubicin-resistant cells. To examine its function in
chemoresistance, RNA interference (RNAi) and forskolin stimulation experiments further demonstrated that ICBP90
and Topo IIα were involved in the proliferation of cells that had acquired doxorubicin resistance. In MCF-7/Dox and
ICBP90-siRNA cells, the cell growth wasn't inhibited by doxorubicin and preferentially arrested in G1 phase. However,
after forskolin increased the Topo IIα expression, these breast cancer cells were again found to be inhibited by
doxorubicin. Further, immunohistochemical assay breast cancer patients accepted EFC regimen showed ICBP90 was
significantly associated with tumor cell proliferation, locally advanced disease and Topo IIα expression. In conclusion,
down-regulation of ICBP90 induced the descended expression of Topo IIα protein which is the target enzyme of
doxorubicin.
~1 Citing
432. A CRE that binds CREB and contributes to PKA-dependent regulation of the proximal promoter of human
RAB25 gene
By Xue Huiping; Qiao Yongxia; Ni Peihua; Wang Jiayi; Chen Changqiang; Huang Gang
MEDLINE
RAB25 plays an important role in tumor progression and aggressiveness; altered RAB25 expression may cause
human cancer. As the underlying mechanism of RAB25-mediated carcinogenesis in various tumor types progressively
comes to light, RAB25 is expected to represent a novel therapeutic target. However, the regulation of RAB25
expression per se has not yet been described. Here we have firstly identified and characterized the human RAB25
promoter. Using PCR-based chromatin accessibility and chromatin immunoprecipitation (ChIP) assays, an open
chromatin conformation (-173/+17) was detected around the transcription start site of the RAB25 gene. Deletion
constructs of the 5' flanking region were fused to a luciferase reporter gene. After transient transfection in gastric
cancer cell line AGS, a CRE (-67/-58) binding CREB was identified in the core promoter region. Electrophoretic
mobility shift (EMSA) and ChIP assays demonstrated that CREB binds to the core promoter. Deletion of CREB
consensus sequence resulted in the total loss of the promoter activity. Moreover, we have also found forskolin, PKA
activator, could enhance open chromatin accessibility, by which to expose the CRE and facilitate phosphorylation of
CREB, which in turn recruits co-factor CBP and Brg I and then results in a more open chromatin configuration
associated with local histone modification, finally heightening RAB25 expression and strengthening its promoter
activity. Therefore, the present study delineates the fundamental elements of a core promoter structure that will be
helpful for future studies regarding the regulation of RAB25 gene.
~5 Citings
433. Impact of gsp oncogene on the mRNA content for somatostatin and dopamine receptors in human
somatotropinomas
By Taboada Giselle Fernandes; Neto Leonardo Vieira; Luque Raul M; Cordoba-Chacon Jose; de Oliveira Machado
Evelyn; de Carvalho Denise Pires; Kineman Rhonda D; Gadelha Monica Roberto
From Neuroendocrinology (2011), 93(1), 40-7, Language: English, Database: MEDLINE
INTRODUCTION: It has been reported in some series that gsp+ somatotropinomas are more sensitive to somatostatin
analogues (SA) and dopamine's actions which may be related to their somatostatin receptor (SSTR) and dopamine
receptor (DR) profile. No previous studies have been undertaken to evaluate the SSTR and DR profile related with the
gsp status in somatotropinomas. OBJECTIVES: To determine if (1) gsp status is correlated with response to
octreotide LAR (LAR) and tumor expression patterns of SSTR1-5 and DR1-5 and (2) cAMP level can directly modulate
SSTR and DR mRNA levels. METHODS: Response to SA was evaluated by GH and IGF-I percent reduction after 3
and 6 months of treatment with LAR. Conventional PCR and sequencing were used to identify gsp+ tumors.
Quantitative real-time PCR was used to determine SSTR and DR tumor expression. Primary pituitary cell cultures of
primates were used to study whether SSTR and DR expression is regulated by forskolin. RESULTS: The response to
LAR did not significantly differ between patients with gsp+ and gsp- tumors; however, gsp+ tumors expressed higher
levels of SSTR1, SSTR2, DR2 and a lower level of SSTR3. Forskolin increased SSTR1, SSTR2, DR1 and DR2
expression in cell cultures. CONCLUSION: Elevated SSTR1, SSTR2, and DR2 tumor expression may help improve
responsiveness to SA and DA therapy; however, this study may not have been appropriately powered to observe
significant effects in the clinical response. Elevated cAMP levels could be directly responsible for the upregulation in
SSTR1, SSTR2 and DR2 mRNA levels observed in gsp+ patients.
~2 Citings
434. Lubiprostone activates Cl- secretion via cAMP signaling and increases membrane CFTR in the human colon
carcinoma cell line, T84
By Ao Mei; Venkatasubramanian Jayashree; Boonkaewwan Chaiwat; Ganesan Nivetha; Syed Asma; Benya Richard
V; Rao Mrinalini C
From Digestive diseases and sciences (2011), 56(2), 339-51, Language: English, Database: MEDLINE
BACKGROUND: Lubiprostone, used clinically (b.i.d.) to treat constipation, has been reported to increase
transepithelial Cl(-) transport in T84 cells by activating ClC-2 channels. AIM: To identify the underlying signaling
pathway, we explored the effects of short-term and overnight lubiprostone treatment on second messenger signaling
and Cl(-) transport. METHODS: Cl(-) transport was assessed either as I(sc) across T84 monolayers grown on
Transwells and mounted in Ussing chambers or by the iodide efflux assay. [cAMP](i) was measured by enzyme
immunoassay, and [Ca(2+)](i) by Fluo-3 fluorescence. Quantitation of apical cell surface CFTR protein levels was
assessed by Western blotting and biotinylation with the EZ-Link Sulfo-NHS-LC-LC-Biotin. ClC-2 mRNA level was
studied by RT-PCR. RESULTS: Lubiprostone and the cAMP stimulator, forskolin, caused comparable and maximal
increases of I(sc) in T84 cells. The I(sc) effects of lubiprostone and forskolin were each suppressed if the tissue had
previously been treated with the other agent. These responses were unaltered even if the monolayers were treated
with lubiprostone overnight. Lubiprostone-induced increases in iodide efflux were ~80% of those obtained with
forskolin. Lubiprostone increased [cAMP](i). H89, bumetanide, or CFTR(inh)-172 greatly attenuated lubiprostone-
stimulated Cl(-) secretion, whereas the ClC-2 inhibitor CdCl(2) did not. Compared to controls, FSK-treatment
increased membrane-associated CFTR by 1.9 fold, and lubiprostone caused a 2.6-fold increase in apical membrane
CFTR as seen by immunoblotting following cell surface biotinylation. CONCLUSIONS: Lubiprostone activates Cl(-)
secretion in T84 cells via cAMP, protein kinase A, and by increasing apical membrane CFTR protein.
~13 Citings
435. Tumor-secreted PGE2 inhibits CCL5 production in activated macrophages through cAMP/PKA signaling
pathway
By Qian Xuesong; Zhang Jidong; Liu Jianguo
One of the major characteristics of tumors is their ability to evade immunosurveillance through altering the properties
and functions of host stromal and/or immune cells. CCL5 has been shown to play important roles in T cell proliferation,
IFN-γ, and IL-2 production, which promotes the differentiation and proliferation of Th1 cells important for immune
defense against intracellular infection. In this study we found that tumor-bearing mice were more susceptible to
bacterial infection and showed reduced CCL5 levels in serum during endotoxic shock. Our data further demonstrated
that the soluble factors secreted by mammary gland tumor cells but not normal mammary gland epithelial cells
inhibited CCL5 expression in macrophages in response to LPS, but not to TNF-α stimulation. The inhibitory effect of
tumor-secreted molecules on LPS-induced CCL5 expression was regulated at the post-transcriptional level. Blocking
PGE(2) synthesis by NS398 or through the use of PGE(2) receptor antagonists AH-6809 (EP2 antagonist) and AH-
23848 (EP4 antagonist) completely reversed the inhibitory effect of tumor-conditioned medium (TCM) on LPS-induced
CCL5 expression. Moreover, PGE(2) and the cAMP analog forskolin could mimic tumor-mediated CCL5 inhibition, and
the inhibitory effects of TCM, PGE(2), and cAMP analog on LPS-induced CCL5 expression could be completely
reversed by the PKA inhibitor H89. Furthermore, blocking PGE(2) synthesis in vivo led to partial recovery of CCL5
production during endotoxic shock. Taken together, our data indicate that PGE(2) secreted from breast cancer cells
suppresses CCL5 secretion in LPS-activated macrophages through a cAMP/PKA signaling pathway, which may result
in suppression of host immune responses against subsequent bacterial infection.
~16 Citings
436. Chronic regulation of colonic epithelial secretory function by activation of G protein-coupled receptors
By Toumi F; Frankson M; Ward J B; Kelly O B; Mroz M S; Bertelsen L S; Keely S J
From Neurogastroenterology and motility : the official journal of the European Gastrointestinal Motility Society (2011),
23(2), 178-86, e43, Language: English, Database: MEDLINE
BACKGROUND: Enteric neurotransmitters that act at G protein-coupled receptors (GPCRs) are well known to acutely
promote epithelial Cl(-) and fluid secretion. Here we examined if acute GPCR activation might have more long-term
consequences for epithelial secretory function. METHODS: Cl(-) secretion was measured as changes in short-circuit
current across voltage-clamped T(84) colonic epithelial cells. Protein expression was measured by western blotting
and intracellular Ca(2+) levels by Fura-2 fluorescence. KEY RESULTS: While acute (15 min) treatment of T(84) cells
with a cholinergic G(q) PCR agonist, carbachol (CCh), rapidly stimulated Cl(-) secretion, subsequent CCh-induced
responses were attenuated in a biphasic manner. The first phase was transient and resolved within 6 h but this was
followed by a chronic phase of attenuated responsiveness that was sustained up to 48 h. CCh-pretreatment did not
chronically alter responses to another G(q)PCR agonist, histamine, or to thapsigargin or forskolin which elevate
intracellular Ca(2+) and cAMP, respectively. This chronically acting antisecretory mechanism is not shared by
neurotransmitters that activate G(s)PCRs. Conditioned medium from CCh-pretreated cells mimicked its chronic
antisecretory actions, suggesting involvement of an epithelial-derived soluble factor but further experimentation ruled
out the involvement of epidermal growth factor receptor ligands. Acute CCh exposure did not chronically alter surface
expression of muscarinic M(3) receptors but inhibited intracellular Ca(2+) mobilization upon subsequent agonist
challenge. CONCLUSIONS & INFERENCES: These data reveal a novel, chronically acting, antisecretory mechanism
that downregulates epithelial secretory capacity upon repeated G(q)PCR agonist exposure. This mechanism involves
release of a soluble factor that uncouples receptor activation from downstream prosecretory signals.
~0 Citings
437. Pituitary adenylyl cyclase activating polypeptide inhibits gli1 gene expression and proliferation in primary
medulloblastoma derived tumorsphere cultures
By Cohen Joseph R; Resnick Daniel Z; Niewiadomski Pawel; Dong Hongmei; Liau Linda M; Waschek James A
From BMC cancer (2010), 10676, Language: English, Database: MEDLINE
BACKGROUND: Hedgehog (HH) signaling is critical for the expansion of granule neuron precursors (GNPs) within the
external granular layer (EGL) during cerebellar development. Aberrant HH signaling within GNPs is thought to give
rise to medulloblastoma (MB) - the most commonly-observed form of malignant pediatric brain tumor. Evidence in both
invertebrates and vertebrates indicates that cyclic AMP-dependent protein kinase A (PKA) antagonizes HH signalling.
Receptors specific for the neuropeptide pituitary adenylyl cyclase activating polypeptide (PACAP, gene name
ADCYAP1) are expressed in GNPs. PACAP has been shown to protect GNPs from apoptosis in vitro, and to interact
with HH signaling to regulate GNP proliferation. PACAP/ptch1 double mutant mice exhibit an increased incidence of
MB compared to ptch1 mice, indicating that PACAP may regulate HH pathway-mediated MB pathogenesis.
METHODS: Primary MB tumorsphere cultures were prepared from thirteen ptch1+/-/p53+/- double mutant mice and
treated with the smoothened (SMO) agonist purmorphamine, the SMO antagonist SANT-1, the neuropeptide PACAP,
the PKA activator forskolin, and the PKA inhibitor H89. Gene expression of gli1 and [3H]-thymidine incorporation were
assessed to determine drug effects on HH pathway activity and proliferation, respectively. PKA activity was
determined in cell extracts by Western blotting using a phospho-PKA substrate antibody. RESULTS: Primary tumor
cells cultured for 1-week under serum-free conditions grew as tumorspheres and were found to express PAC1 receptor
transcripts. Gli1 gene expression was significantly reduced by SANT-1, PACAP and forskolin, but was unaffected by
purmorphamine. The attenuation of gli1 gene expression by PACAP was reversed by the PKA inhibitor H89, which
also blocked PKA activation. Treatment of tumorsphere cultures with PACAP, forskolin, and SANT-1 for 24 or 48
hours reduced proliferation. CONCLUSIONS: Primary tumorspheres derived from ptch1+/-/p53+/- mice exhibit
constitutive HH pathway activity. PACAP antagonizes HH signalling in these cells in a manner blocked by the PKA
antagonist H89. PACAP and pharmacological activation of PKA also inhibited proliferation. Our data suggests that
regulation of HH signaling by PACAP/PKA signaling may provide an alternative to SMO inhibition for the treatment of
MB.
~9 Citings
438. Frequent phosphodiesterase 11A gene (PDE11A) defects in patients with Carney complex (CNC) caused by
PRKAR1A mutations: PDE11A may contribute to adrenal and testicular tumors in CNC as a modifier of the
phenotype
By Libe Rossella; Horvath Anelia; Vezzosi Delphine; Fratticci Amato; Coste Joel; Perlemoine Karine; Ragazzon Bruno;
Guillaud-Bataille Marine; Groussin Lionel; Clauser Eric; et al
MEDLINE
BACKGROUND: Carney complex (CNC) is an autosomal dominant multiple neoplasia, caused mostly by inactivating
mutations of the regulatory subunit 1A of the protein kinase A (PRKAR1A). Primary pigmented nodular adrenocortical
disease (PPNAD) is the most frequent endocrine manifestation of CNC with a great inter-individual variability.
Germline, protein-truncating mutations of phosphodiesterase type 11A (PDE11A) have been described to predispose
to a variety of endocrine tumors, including adrenal and testicular tumors. OBJECTIVES: Our objective was to
investigate the role of PDE11A as a possible gene modifier of the phenotype in a series of 150 patients with CNC.
RESULTS: A higher frequency of PDE11A variants in patients with CNC compared with healthy controls was found
(25.3 vs. 6.8%, P < 0.0001). Among CNC patients, those with PPNAD were significantly more frequently carriers of
PDE11A variants compared with patients without PPNAD (30.8 vs. 13%, P = 0.025). Furthermore, men with PPNAD
were significantly more frequently carriers of PDE11A sequence variants (40.7%) than women with PPNAD (27.3%) (P
< 0.001). A higher frequency of PDE11A sequence variants was also found in patients with large-cell calcifying Sertoli
cell tumors (LCCSCT) compared with those without LCCSCT (50 vs. 10%, P = 0.0056). PDE11A variants were
significantly associated with the copresence of PPNAD and LCCSCT in men: 81 vs. 20%, P < 0.004). The
simultaneous inactivation of PRKAR1A and PDE11A by small inhibitory RNA led to an increase in cAMP-regulatory
element-mediated transcriptional activity under basal conditions and after stimulation by forskolin. CONCLUSIONS:
We demonstrate, in a large cohort of CNC patients, a high frequency of PDE11A variants, suggesting that PDE11A is a
genetic modifying factor for the development of testicular and adrenal tumors in patients with germline PRKAR1A
mutation.
~17 Citings
439. Signaling from the human melanocortin 1 receptor to ERK1 and ERK2 mitogen-activated protein kinases
involves transactivation of cKIT
By Herraiz Cecilia; Journe Fabrice; Abdel-Malek Zalfa; Ghanem Ghanem; Jimenez-Cervantes Celia; Garcia-Borron
Jose C
From Molecular endocrinology (Baltimore, Md.) (2011), 25(1), 138-56, Language: English, Database: MEDLINE
Melanocortin 1 receptor (MC1R), a Gs protein-coupled receptor expressed in melanocytes, is a major determinant of
skin pigmentation, phototype and cancer risk. Upon stimulation by αMSH, MC1R triggers the cAMP and ERK1/ERK2
MAPK pathways. In mouse melanocytes, ERK activation by αMSH binding to Mc1r depends on cAMP, and
melanocytes are considered a paradigm for cAMP-dependent ERK activation. However, human MC1R variants
associated with red hair, fair skin [red hair color (RHC) phenotype], and increased skin cancer risk display reduced
cAMP signaling but activate ERKs as efficiently as wild type in heterologous cells, suggesting independent signaling to
ERKs and cAMP in human melanocytes. We show that MC1R signaling activated the ERK pathway in normal human
melanocytes and melanoma cells expressing physiological levels of endogenous RHC variants. ERK activation was
comparable for wild-type and mutant MC1R and was independent on cAMP because it was neither triggered by
stimulation of cAMP synthesis with forskolin nor blocked by the adenylyl cyclase inhibitor 2',5'-dideoxyadenosine.
Stimulation of MC1R with αMSH did not lead to protein kinase C activation and ERK activation was unaffected by
protein kinase C inhibitors. Conversely, pharmacological interference, small interfering RNA studies, expression
profiles, and functional reconstitution experiments showed that αMSH-induced ERK activation resulted from Src
tyrosine kinase-mediated transactivation of the stem cell factor receptor, a receptor tyrosine kinase essential for
proliferation, differentiation, and survival of melanocyte precursors, thus demonstrating a functional link between the
stem cell factor receptor and MC1R. Moreover, this transactivation phenomenon is unique because it is unaffected by
natural mutations impairing canonical MC1R signaling through the cAMP pathway.
~18 Citings
440. cAMP and fibroblast growth factor 2 regulate bone sialoprotein gene expression in human prostate cancer
cells
By Li Zhengyang; Sasaki Yoko; Mezawa Masaru; Wang Shuang; Li Xinyue; Yang Li; Wang Zhitao; Zhou Liming; Araki
Shouta; Matsumura Hiroyoshi; et al
From Gene (2011), 471(1-2), 1-12, Language: English, Database: MEDLINE
Bone sialoprotein (BSP) is a noncollagenous protein of the extracellular matrix in mineralized connective tissues that
has been implicated in the nucleation of hydroxyapatite. Forskolin (FSK), an activator of adenylate cyclase, increased
the intracellular cAMP level, which stimulates the proliferation and differentiation of osteoblasts. Fibroblast growth
factor 2 (FGF2) is a potent mitogen in many cell types, including osteoblasts. In human prostate cancer DU145 cells,
FSK (1 µM) and FGF2 (10 ng/ml) increased BSP and Runx2 mRNA and protein levels at 3 and 12h, respectively.
Transient transfection analyses were performed using chimeric constructs of the human BSP gene promoter linked to a
luciferase reporter gene. Treatment of DU145 cells with FSK (1 µM) and FGF2 (10 ng/ml) increased the luciferase
activities of constructs between -60LUC to -927LUC and -108LUC to -927LUC, including the human BSP gene
promoter. Effects of FSK and FGF2 abrogated in constructs included 2bp mutations in the two cAMP response
elements (CRE1 and CRE2). Luciferase activities induced by FSK and FGF2 were blocked by protein kinase A and
tyrosine kinase inhibitors. Gel mobility shift analyses showed that FSK and FGF2 increased the binding of CRE1 and
CRE2. CRE1-protein complexes were supershifted by phospho-CREB1 and c-Fos antibodies, and disrupted by
CREB1, c-Jun, JunD, Fra2, p300, Runx2, Dlx5 and Smad1 antibodies. CRE2-protein complexes were disrupted by
CREB1, phospho-CREB1, c-Fos, c-Jun, JunD, Fra2, p300, Runx2, Dlx5 and Smad1 antibodies. These studies
demonstrate that FSK and FGF2 stimulate BSP transcription in DU145 human prostate cancer cells by targeting the
CRE1 and CRE2 elements in the human BSP gene promoter.
~2 Citings
441. Multiple roles of protein kinase a in arachidonic acid-mediated Ca2+ entry and tumor-derived human
endothelial cell migration
By Fiorio Pla Alessandra; Genova Tullio; Pupo Emanuela; Tomatis Cristiana; Genazzani Armando; Zaninetti Roberta;
Munaron Luca
We recently showed that arachidonic acid (AA) triggers calcium signals in endothelial cells derived from human breast
carcinoma (B-TEC). In particular, AA-dependent Ca(2+) entry is involved in the early steps of tumor angiogenesis in
vitro. Here, we investigated the multiple roles of the nitric oxide (NO) and cyclic AMP/protein kinase A (PKA) pathways
in AA-mediated Ca(2+) signaling in the same cells. B-TEC stimulation with 5 µmol/L AA resulted in endothelial NO
synthase (NOS) phosphorylation at Ser(1177), and NO release was measured with the fluorescent NO-sensitive probe
DAR4M-AM. PKA inhibition by the use of the membrane-permeable PKA inhibitory peptide myristoylated PKI(14-22)
completely prevented both AA- and NO-induced calcium entry and abolished B-TEC migration promoted by AA. AA-
dependent calcium entry and cell migration were significantly affected by both the NOS inhibitor N(G)-nitro-l-arginine
methyl ester and the NO scavenger 2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl 3-oxide, suggesting that NO
release is functionally involved in the signaling dependent on AA. Moreover, pretreatment with carboxyamidotriazole,
an antiangiogenic compound that interferes with agonist-activated calcium entry, prevented AA-dependent B-TEC
motility. Interestingly, even in the absence of AA, enhancement of the cyclic AMP/PKA pathway with the adenylyl
cyclase activator forskolin evoked a calcium entry dependent on NOS recruitment and NO release. The functional
relevance of AA-induced calcium entry could be restricted to tumor-derived endothelial cells (EC) because AA evoked
a smaller calcium entry in normal human microvascular ECs compared with B-TECs, and even more importantly, it was
unable to promote cell motility in wound healing assay. This evidence opens an intriguing opportunity for differential
pharmacologic treatment between normal and tumor-derived human ECs.
~7 Citings
442. Adiponectin promotes syncytialisation of BeWo cell line and primary trophoblast cells
By Benaitreau Delphine; Dos Santos Esther; Leneveu Marie-Christine; De Mazancourt Philippe; Pecquery Rene;
Dieudonne Marie-Noelle
From Reproductive biology and endocrinology : RB&E (2010), 8128, Language: English, Database: MEDLINE
BACKGROUND: In human pregnancy, a correct placentation depends on trophoblast proliferation, differentiation,
migration and invasion. These processes are highly regulated by placental hormones, growth factors and cytokines.
Recently, we have shown that adiponectin, an adipokine, has anti-proliferative effects on trophoblastic cells. Here, we
complete this study by demonstrating that adiponectin modulates BeWo and human villous cytotrophoblast cell
differentiation. RESULTS: We showed that hCG secretion was up-regulated by adiponectin treatment in both BeWo
cells and human cytotrophoblasts from very early placentas (5-6 weeks). The expression of two trophoblast
differentiation markers, leptin and syncytin 2, was also up-regulated by adiponectin in BeWo cells. Moreover,
adiponectin treatment induced a loss of E-cadherin staining in these cells. In parallel, we demonstrated that AdipoR1
and AdipoR2 are up-regulated during forskolin induced BeWo cell differentiation, reinforcing the role of adiponectin in
trophoblast syncytialization. SiRNA mediated down-regulation of AdipoR1 and AdipoR2 was used to demonstrate that
adiponectin effects on differentiation were essentially mediated by these receptors. Finally, using a specific inhibitor,
we demonstrated that the PKA signalling pathway could be one pathway involved in adiponectin effects on trophoblast
differentiation. CONCLUSION: Adiponectin enhances the differentiation process of trophoblast cells and could thus
be involved in functional syncytiotrophoblast formation.
~7 Citings
443. The effect of valproate and levetiracetam on steroidogenesis in forskolin-stimulated H295R cells
By von Krogh Kristine; Harjen Hannah; Almas Camilla; Zimmer Karin E; Dahl Ellen; Olsaker Ingrid; Tauboll Erik;
Ropstad Erik; Verhaegen Steven
From Epilepsia (2010), 51(11), 2280-8, Language: English, Database: MEDLINE
PURPOSE: Endocrine disruptive effects have been frequently observed in patients using antiepileptic drugs (AEDs).
Two different AEDs, valproate (VPA) and levetiracetam (LEV), were tested in forskolin-stimulated human adrenal
carcinoma (H295R) cells to explore their effect on steroidogenesis. VPA has a long history as an anticonvulsant and is
linked with many of the endocrine disorders associated with AED use. LEV is a newer AED, and no endocrine
disruptive effects have been reported in humans to date. METHODS: H295R cells, which are capable of full
steroidogenesis, were stimulated with forskolin and exposed to either VPA or LEV for 48 h. Medium was collected and
analyzed for hormone production. For the VPA-exposed cells, steroidogenic gene expression analysis was also
conducted. RESULTS: VPA exposure resulted in a significant reduction in progesterone and estradiol (E2)
production, whereas testosterone (T) levels remained unchanged. There were also significant alterations in expression
level for most genes analyzed. LEV exposure resulted in a minor, but statistically significant, reduction in T and E2
production. DISCUSSION: Exposure of forskolin-stimulated H295R cells to VPA led to an increased T/E2 ratio
through a significant decrease in estradiol production. Gene analysis suggested that VPA affects NR0B1 expression.
NR0B1 inhibits promoters of other genes involved in steroidogenesis, and the altered expression of NR0B1 might
explain the observed down-regulation in hormone production. The effects of LEV exposure on hormone secretion
were not considered to be biologically significant.
~1 Citing
444. Evaluation of a bioluminescent mouse model expressing aromatase PII-promoter-controlled luciferase as a

tool for the study of endocrine disrupting chemicals
By Rivest Patricia; Devine Patrick J; Sanderson J Thomas
From Toxicology and applied pharmacology (2010), 249(1), 33-40, Language: English, Database: MEDLINE
Dysfunction of the enzyme aromatase (CYP19) is associated with endocrine pathologies such as osteoporosis,
impaired fertility and development of hormone-dependent cancers. Certain endocrine disrupting chemicals affect
aromatase expression and activity in vitro, but little is known about their ability to do so in vivo. We evaluated a
bioluminescent mouse model (LPTA®)CD-1-Tg(Cyp19-luc)-Xen) expressing luciferase under control of the gonadal
aromatase pII promoter as an in vivo screening tool for chemicals that may affect aromatase expression. We studied
the effects of forskolin, pregnant mare serum gonadotropin and atrazine in this model (atrazine was previously shown
to induced pII-promoter-driven aromatase expression in H295R human adrenocortical carcinoma cells). About 2-4 out
of every group of 10 male or female Cyp19-luc mice injected i.p. with 10 mg/kg forskolin had increased gonadal
bioluminescence after 3-5 days compared to controls; the others appeared non-responsive. Similarly, about 4 per
group of 9 individual females injected with pregnant mare serum gonadotropin had increased ovarian bioluminescence
after 24 h. There was a statistically significant correlation between ovarian bioluminescence and plasma estradiol
concentrations (n=14; p=0.022). Males exposed to a single dose of 100 mg/kg or males and females exposed to 5
daily injections of 30 mg/kg atrazine showed no change in gonadal bioluminescence over a 7 day period, but a
significant interaction was found between atrazine (100 mg/kg) and time in female mice (p<0.05; two-way ANOVA). Ex
vivo luciferase activity in dissected organs was increased by forskolin in testis, epididymis and ovaries. Atrazine (30
mg/kg/day) increased (30%) luciferase activity significantly in epididymis only. In conclusion, certain individual Cyp19-
luc mice are highly responsive to aromatase inducers, suggesting this model, with further optimization, may have
potential as an in vivo screening tool for environmental contaminants.
~1 Citing
445. CFTR and calcium-activated chloride channels in primary cultures of human airway gland cells of serous or
mucous phenotype
By Fischer Horst; Illek Beate; Sachs Lorne; Finkbeiner Walter E; Widdicombe Jonathan H
From American journal of physiology. Lung cellular and molecular physiology (2010), 299(4), L585-94, Language:
English, Database: MEDLINE
Using cell culture models, we have investigated the relative importance of cystic fibrosis transmembrane conductance
regulator (CFTR) and calcium-activated chloride channels (CaCC) in Cl secretion by mucous and serous cells of
human airway glands. In transepithelial recordings in Ussing chambers, the CFTR inhibitor CFTR(inh)-172 abolished
60% of baseline Cl secretion in serous cells and 70% in mucous. Flufenamic acid (FFA), an inhibitor of CaCC,
reduced baseline Cl secretion by ∼20% in both cell types. Methacholine and ATP stimulated Cl secretion in both cell
types, which was largely blocked by treatment with 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (BAPTA)
and partially by mucosal FFA or CFTR(inh)-172 with the exception of methacholine responses in mucous cells, which
were not blocked by FFA and partially (∼60%) by CFTR(inh)-172. The effects of ionomycin on short-circuit current
(I(sc)) were less than those of ATP or methacholine. Forskolin stimulated Cl secretion only if Cl in the mucosal
medium was replaced by gluconate. In whole cell patch-clamp studies of single isolated cells, cAMP-induced Cl
currents were ∼3-fold greater in serous than mucous cells. Ionomycin-induced Cl currents were 13 times (serous) or
26 times (mucous) greater than those generated by cAMP and were blocked by FFA. In serous cells, mRNA for
transmembrane protein 16A (TMEM16A) was ∼10 times more abundant than mRNA for CFTR. In mucous cells it was
∼100 times more abundant. We conclude: 1) serous and mucous cells both make significant contributions to gland
fluid secretion; 2) baseline Cl secretion in both cell types is mediated predominantly by CFTR, but CaCC becomes
increasingly important after mediator-induced elevations of intracellular Ca; and 3) the high CaCC currents seen in
patch-clamp studies and the high TMEM16A expression in intact polarized cells sheets are not reflected in
transepithelial current recordings.
~8 Citings
446. Leptin potentiates antiproliferative action of cAMP elevation via protein kinase A down-regulation in breast
cancer cells
By Naviglio Silvio; Di Gesto Davide; Illiano Fausto; Chiosi Emilio; Giordano Antonio; Illiano Gennaro; Spina Annamaria
Previously, we have shown that leptin potentiates the antiproliferative action of cAMP elevating agents in breast cancer
cells and that the protein kinase A (PKA) inhibitor KT-5720 prevented the antiproliferative effects induced by the leptin
plus cAMP elevation. The present experiments were designed to gain a better understanding about the PKA role in
the antitumor interaction between leptin and cAMP elevating agents and on the underlying signaling pathways. Here
we show that exposure of MDA-MB-231 breast cancer cells to leptin resulted in a strong phosphorylation of both
ERK1/2 and STAT3. Interestingly, intracellular cAMP elevation upon forskolin pretreatment completely abrogated both
ERK1/2 and STAT3 phosphorylation in response to leptin and was accompanied by a consistent CREB
phosphorylation. Notably, leptin plus forskolin cotreatments resulted in a strong decrease of both PKA regulatory RIα
and catalytic subunits protein levels. Importantly, pretreatment with the PKA inhibitor KT-5720 blocked the forskolin-
induced CREB phosphorylation and prevented both the inhibition by forskolin of leptin-induced ERK1/2 and STAT3
phosphorylation and the PKA subunits down-regulation induced by the combination of leptin and forskolin. Altogether,
our results indicate that leptin-dependent signaling pathways are influenced by cAMP elevation and identify PKA as
relevantly involved in the pharmacological antitumor interaction between leptin and cAMP elevating drugs in MDA-MB-
231 cells. We propose a molecular model by which PKA confers its effects. Potential therapeutic applications by our
data will be discussed.
~8 Citings
447. Cyclin D1 and cyclin D3 show divergent responses to distinct mitogenic stimulation
By Anderson Alexandra A; Child Emma S; Prasad Aarathi; Elphick Lucy M; Mann David J
D-type cyclins predominantly regulate progression through the cell cycle by their interactions with cyclin-dependent
kinases (cdks). Here, we show that stimulating mitogenesis of Swiss 3T3 cells with phorbol esters or forskolin can
induce divergent responses in the expression levels, localization and activation state of cyclin D1 and cyclin D3.
Phorbol ester-mediated protein kinase C stimulation induces S phase entry which is dependent on MAPK activation
and increases the levels and activation of cyclin D1, whereas forskolin-mediated cAMP-dependent protein kinase A
stimulation induces mitogenesis that is independent of MAPK, but dependent upon mTor and specifically increases the
level and activation of cyclin D3. These findings uncover additional levels of complexity in the regulation of the cell
cycle at the level of the D-type cyclins and thus may have important therapeutic implications in cancers where specific
D-cyclins are overexpressed.
~0 Citings
448. Sunitinib inhibits MEK/ERK and SAPK/JNK pathways and increases sodium/iodide symporter expression in
papillary thyroid cancer
By Fenton Mike S; Marion Kenneth M; Salem Andrew K; Hogen Rachel; Naeim Faramarz; Hershman Jerome M
From Thyroid : official journal of the American Thyroid Association (2010), 20(9), 965-74, Language: English,
Database: MEDLINE
BACKGROUND: Sunitinib malate (Sutent, Pfizer, Inc.; SU11248) is a selective, multitargeted inhibitor of receptor
tyrosine kinases and has been shown to inhibit receptors for VEGF, PDGF, KIT, FLT3, and RET. The objective of this
study was to determine the effects of sunitinib on signal transduction pathways and on gene expression of iodide-
metabolizing proteins in papillary cancer cells with the RET/PTC1 rearrangement. METHODS: We investigated the
effects of sunitinib on cell growth, signal transduction pathways, and thyroid-specific gene expression in papillary
thyroid cancer (PTC) cell lines that had the RET/PTC1 rearrangement. RESULTS: Sunitinib inhibited proliferation of
RET/PTC1 subclones in a time- and dose-related manner. The mean 50% lethal concentration in the RET/PTC1
subclones was 1.81 microM. Incubation of RET/PTC1 cells with 1 microM sunitinib inhibited their migration potential
and transformed their morphology. Sunitinib inhibited RET autophosphorylation at Y1062 and the activation of signal
transducer and activator of transcription 3 by blocking Y705 phosphorylation. Sunitinib caused cell cycle arrest in the
G0/G1 phase and dephosphorylation of retinoblastoma protein, but did not induce apoptosis. Western blot analysis of
the p38, MEK/ERK, and SAPK/JNK mitogen-activated protein kinase signal transduction pathways showed that
sunitinib blocked ERK 1/2 and JNK phosphorylation in the cytoplasm. Sunitinib treatment of RET/PTC1 cell lines, in
combination, with forskolin induced expression of the sodium (Na)/iodide (I) symporter (NIS) and the transcription
factors that bind the NIS upstream enhancer. Mechanistically, the inhibition of both MEK/ERK and SAPK/JNK
cytoplasmic pathways individually and in combination caused an increase in NIS gene expression. CONCLUSION:
Sunitinib appears to target the cytosolic MEK/ERK and SAPK/JNK pathways in the RET/PTC1 cell lines, suggesting
that blocking these pathways is at least part of the mechanism by which sunitinib inhibits cell proliferation and causes
stimulation of NIS gene expression in RET/PTC1 cells.
~6 Citings
449. Expression of cAMP-dependent protein kinase isoforms in the human prostate: functional significance and
relation to PDE4
By Waldkirch Eginhard; Uckert Stefan; Sigl Katja; Langnaese Kristina; Richter Karin; Stief Christian G; Kuczyk Markus
A; Hedlund Petter
From Urology (2010), 76(2), 515.e8-14, Language: English, Database: MEDLINE
OBJECTIVES: To investigate the expression of isoforms of the cyclic AMP (cAMP)-dependent protein kinase (cAK) in
the transition zone of the human prostate and the functional significance of the enzyme in the control of prostate
smooth muscle. METHODS: Using Western blot analysis and immunohistochemistry, the expression and distribution
in the prostate of cAKIalpha, cAKIbeta, cAKIIalpha, and cAKIIbeta in relation to alpha-actin and the phosphodiesterase
PDE4 (types A and B) were investigated. The effects of the cAK inhibitor Rp-8-CPT-cAMPS on the reversion of the
adrenergic tension of isolated prostate tissue induced by forskolin, rolipram, sodium nitroprusside (SNP), and tadalafil
were examined by means of the organ bath technique. RESULTS: Immunosignals specific for cAKIalpha, cAKIIalpha,
and cAKIIbeta were observed in the smooth musculature and glandular structures of the prostate. Double stainings
revealed the colocalization of alpha-actin and PDE4 with the cAK isoforms. The expression of the cAK isoforms was
confirmed by Western blot analysis. The relaxation of the tension induced by norepinephrine brought about by
forskolin, rolipram, SNP, and tadalafil was significantly attenuated by Rp-8-CPT-cAMPS. CONCLUSIONS: The
colocalization of smooth muscle alpha-actin and PDE4 with cAK, as well as the results from the organ bath
experiments, provide further evidence for a pivotal role of the cAMP-dependent signaling in the regulation of prostate
smooth muscle contractility. Compounds interacting with the cAMP/cAK pathway might represent a new therapeutic
avenue to treat symptoms of benign prostatic hyperplasia and lower urinary tract symptomatology.
~6 Citings
450. Metformin inhibits aromatase expression in human breast adipose stromal cells via stimulation of AMP-
activated protein kinase
By Brown Kristy A; Hunger Nicole I; Docanto Maria; Simpson Evan R
AMP-activated protein kinase (AMPK) is recognized as a master regulator of energy homeostasis. In concert with the
AMPK-kinase LKB1, it has been shown to provide a molecular link between obesity and postmenopausal breast
cancer via its actions to inhibit aromatase expression, hence estrogen production, within the breast. The anti-diabetic
drug metformin is known to increase the activity of AMPK and was therefore hypothesized to inhibit aromatase
expression in primary human breast adipose stromal cells. Results demonstrate that metformin significantly decreases
the forskolin/phorbol ester (FSK/PMA)-induced expression of aromatase at concentrations of 10 and 50 muM.
Consistent with the hypothesized actions of metformin to increase AMPK activity, treatment with 50 muM metformin
results in a significant increase in phosphorylation of AMPK at Thr172. Interestingly, metformin also causes a
significant increase in LKB1 protein expression and promoter activity, thereby providing for the first time an additional
mechanism by which metformin activates AMPK. Furthermore, metformin inhibits the nuclear translocation of CRTC2,
a CREB-coactivator known to increase aromatase expression which is also a direct downstream target of AMPK.
Overall, these results suggest that metformin would reduce the local production of estrogens within the breast thereby
providing a new key therapeutic tool that could be used in the neoadjuvant and adjuvant settings and conceivably also
as a preventative measure in obese women.
~23 Citings
451. Rapid activation of dormant presynaptic terminals by phorbol esters

By Chang Chun Yun; Jiang Xiaoping; Moulder Krista L; Mennerick Steven
From The Journal of neuroscience : the official journal of the Society for Neuroscience (2010), 30(30), 10048-60,
Presynaptic stimulation stochastically recruits transmission according to the release probability (P(r)) of synapses. The
majority of central synapses have relatively low P(r), which includes synapses that are completely quiescent
presynaptically. The presence of presynaptically dormant versus active terminals presumably increases synaptic
malleability when conditions demand synaptic strengthening or weakening, perhaps by triggering second messenger
signals. However, whether modulator-mediated potentiation involves recruitment of transmission from dormant
terminals remains unclear. Here, by combining electrophysiological and fluorescence imaging approaches, we
uncovered rapid presynaptic awakening by select synaptic modulators. A phorbol ester phorbol 12,13-dibutyrate
(PDBu) (a diacylglycerol analog), but not forskolin (an adenylyl cyclase activator) or elevated extracellular calcium,
recruited neurotransmission from presynaptically dormant synapses. This effect was not dependent on protein kinase
C activation. After PDBu-induced awakening, these previously dormant terminals had a synaptic P(r) spectrum similar
to basally active synapses naive to PDBu treatment. Dormant terminals did not seem to have properties of nascent or
immature synapses, judged by NR2B NMDAR (NMDA receptor) receptor subunit contribution after PDBu-stimulated
awakening. Strikingly, synapses rendered inactive by prolonged depolarization, unlike basally dormant synapses,
were not awakened by PDBu. These results suggest that the initial release competence of synapses can dictate the
acute response to second messenger modulation, and the results suggest multiple pathways to presynaptic dormancy
and awakening.
~6 Citings
452. Proteomic analysis reveals ATP-dependent steps and chaperones involvement in luteolin-induced lung
cancer CH27 cell apoptosis
By Lee Hong-Zin; Yang Wen-Hui; Bao Bo-Ying; Lo Pei-Ling
The present study applied 2D electrophoresis to analyze the proteins involved in luteolin (50 microM)-induced CH27
cell apoptosis. We found 7 proteins to be markedly changed. According to the data of analysis of these protein spots,
we hypothesized that ATP synthetic pathway and heat shock proteins were involved in luteolin-induced CH27 cell
apoptosis. In this study, luteolin induced a significant change in intracellular ATP levels and mitochondrial activity of
CH27 cells. Further experiments demonstrated that pretreatment with forskolin blocked the luteolin-induced cell death.
P38 and heat shock protein 27 may be important participants in the luteolin-induced changes in organization of actin
microfilaments in this study. In addition, endoplasmic reticulum stress is also important in the luteolin-induced CH27
cell apoptosis. Our findings suggested that the function of mitochondria and endoplasmic reticulum is the integral
factor in luteolin-induced CH27 cell apoptosis.
~2 Citings
453. Estrogen and non-genomic upregulation of voltage-gated Na(+) channel activity in MDA-MB-231 human
breast cancer cells: role in adhesion
By Fraser Scott P; Ozerlat-Gunduz Iley; Onkal Rustem; Diss James K J; Latchman David S; Djamgoz Mustafa B A
External (but not internal) application of beta-estradiol (E2) increased the current amplitude of voltage-gated Na(+)
channels (VGSCs) in MDA-MB-231 human breast cancer (BCa) cells. The G-protein activator GTP-gamma-S, by
itself, also increased the VGSC current whilst the G-protein inhibitor GDP-beta-S decreased the effect of E2.
Expression of GPR30 (a G-protein-coupled estrogen receptor) in MDA-MB-231 cells was confirmed by PCR, Western
blot and immunocytochemistry. Importantly, G-1, a specific agonist for GPR30, also increased the VGSC current
amplitude in a dose-dependent manner. Transfection and siRNA-silencing of GPR30 expression resulted in
corresponding changes in GPR30 protein expression but only internally, and the response to E2 was not affected. The
protein kinase A inhibitor, PKI, abolished the effect of E2, whilst forskolin, an adenylate cyclase activator, by itself,
increased VGSC activity. On the other hand, pre-incubation of the MDA-MB-231 cells with brefeldin A (a trans-Golgi
protein trafficking inhibitor) had no effect on the E2-induced increase in VGSC amplitude, indicating that such
trafficking ('externalisation') of VGSC was not involved. Finally, acute application of E2 decreased cell adhesion whilst
the specific VGSC blocker tetrodotoxin increased it. Co-application of E2 and tetrodotoxin inhibited the effect of E2 on
cell adhesion, suggesting that the effect of E2 was mainly through VGSC activity. Pre-treatment of the cells with PKI
abolished the effect of E2 on adhesion, consistent with the proposed role of PKA. Potential implications of the E2-
induced non-genomic upregulation of VGSC activity for BCa progression are discussed.
~8 Citings

454. Assessment of chemical effects on aromatase activity using the H295R cell line
By Higley Eric B; Newsted John L; Zhang Xiaowei; Giesy John P; Hecker Markus
From Environmental science and pollution research international (2010), 17(5), 1137-48, Language: English,
Database: MEDLINE
BACKGROUND, AIM, AND SCOPE: In response to concerns about chemical substances that can alter the function of
endocrine systems and may result in adverse effects on human and ecosystem health, a number of in vitro tests have
been developed to identify and assess the endocrine disrupting potential of chemicals and environmental samples.
One endpoint that is frequently used in in vitro models for the assessment of chemical effects on the endocrine system
is the alteration of aromatase activity (AA). Aromatase is the enzyme responsible for converting androgens to
estrogens. Some commonly used aromatase assays, including the human microsomal assay that is a mandatory test
in US-EPA's endocrine disruptor screening program (EDSP), detect only direct effects of chemicals on aromatase
activity and not indirect effects, including changes in gene expression or transcription factors. This can be a problem
for chemical screening initiatives such as the EDSP because chemicals can affect aromatase both indirectly and
directly. Here we compare direct, indirect, and combined measurements of AA using the H295R cell line after
exposure to seven model chemicals. Furthermore, we compare the predictability of the different types of AA
measurements for 17beta-estradiol (E2) and testosterone (T) production in vitro. MATERIALS AND METHODS:
H295R cells were exposed to forskolin, atrazine, letrozole, prochloraz, ketoconazole, aminoglutethimide, and prometon
for 48 h. Direct, indirect, and combined effects on aromatase activity were measured using a tritiated water-release
assay. Direct effects on aromatase activity were assessed by exposing cells only during the conduct of the tritium-
release assay. Indirect effects were measured after exposing cells for 48 h to test chemicals, and then measuring AA
without further chemical addition. Combined AA was measured by exposing cells prior and during the conduction of
the tritium-release assay. Estradiol and testosterone were measured by ELISA. RESULTS AND DISCUSSION:
Exposure to the aromatase inhibitors letrozole, prochloraz, ketoconazole, and aminoglutethimide resulted in greater
indirect aromatase activity after a 48-h exposure due to presumed compensatory mechanisms involved in aromatase
activity regulation. Forskolin and atrazine caused similar changes in hormone production and enzyme profiles, and
both chemicals resulted in a dose-dependent increase in E2, T, and indirect AA. Neither of these two chemicals
directly affected AA. For most of the chemicals, direct and combined AA and E2 were good predictors of the
mechanism of action of the chemical, with regard to AA. Indirect aromatase activity was a less precise predictor of
effects at the hormone level because of presumed feedback loops that made it difficult to predict the chemicals' true
effects, mostly seen with the aromatase inhibitors. Further, it was found that direct and indirect AA measurements
were not reliable predictors of effects on E2 for general inducers and inhibitors, respectively. CONCLUSIONS:
Differential modulation of AA and hormone production was observed in H295R cells after exposure to seven model
chemicals, illustrating the importance of measuring multiple endpoints when describing mechanisms of action in vitro.
RECOMMENDATIONS AND PERSPECTIVES: For future work with the H295R, it is recommended that a combination
of direct and indirect aromatase measurements is used because it was best in predicting the effects of a chemical on
E2 production and its mechanism of action. Further, it was shown that direct AA measurements, which are a common
way to measure AA, must be used with caution in vitro.
~1 Citing
455. Differential regulation and roles of urocortins in human adrenal H295R cells
By Kageyama Kazunori; Hanada Komaki; Suda Toshihiro
From Regulatory peptides (2010), 162(1-3), 18-25, Language: English, Database: MEDLINE
Three urocortins (Ucns) are known as members of the corticotropin-releasing factor (CRF) family of peptides and serve
as natural ligands for CRF receptors. Ucn1 and Ucn3 exhibit potent effects on the adrenal system via the CRF
receptors. This study aimed to explore the regulation and roles of Ucns in the adrenal system using human adrenal
carcinoma H295R cells, which express Ucn1, Ucn2, Ucn3, CRF receptor type 1 (CRF(1) receptor), and CRF receptor
type 2a (CRF(2a) receptor) mRNA. Forskolin, which stimulates adenylate cyclase and then increases intracellular
cAMP production, was shown to transiently decrease Ucn1 and Ucn2 mRNA levels, but increase Ucns 1-3 mRNA
levels in H295R cells. Steroidogenic acute regulatory protein, Cyp11beta1, and Cyp11beta2 mRNA levels, and both
cortisol and aldosterone secretions were elevated by Ucn1. Cell viability was reduced by both Ucn1 and Ucn3 via the
CRF(2) receptor in H295R cells. Ucn1 and Ucn3 increased the expression of the cAMP-response element binding
protein and extracellular signal-related kinase (ERK) phosphorylations. The ERK and protein kinase A pathways were
involved in Ucn3-decreased cell viability.
~2 Citings
456. Effects of catechin, epicatechin and epigallocatechin gallate on testosterone production in rat leydig cells
By Yu Po-Ling; Pu Hsiao-Fung; Chen Sung-Yun; Wang Shyi-Wu; Wang Paulus S
From Journal of cellular biochemistry (2010), 110(2), 333-42, Language: English, Database: MEDLINE
Catechins have been reported to have many pharmacological properties such as the effects of anti-oxidative, anti-
inflammatory, anti-carcinogenic, anti-ultraviolet, and reduction of blood pressure as well as glucose and cholesterol
levels. However, the effect of catechins on the reproductive mechanism is still unknown. In the present study, the
effects of catechins on testosterone secretion in rat testicular Leydig cells (LCs) were explored. Both in vivo and in
vitro investigations were performed. Purified LCs were incubated with or without catechin (CCN), epicatechin (EC),
epigallocatechin gallate (EGCG, 10(-10)-10(-8) M) under challenge with human chorionic gonadotropin (hCG, 0.01
IU/ml), forskolin, SQ22536 (an adenylyl cyclase inhibitor), 8-bromo-adenosine 3':5'-cyclic monophosphate (8-Br-
cAMP), A23187 (a calcium ionophore), and nifedipine (10(-5) M), respectively. To study the effects of catechins on
steroidogenesis, steroidogenic precursors-stimulated testosterone release was examined. The functions of the
steroidogenic enzymes including protein expression of cytochrome P450 side chain cleavage enzyme (P450scc) and
steroidogenic acute regulatory (StAR) protein were investigated and expressed by Western blotting. Catechins
increased plasma testosterone in vivo in male rats. In vitro, low-dose concentration of catechins increased
gonadotropin releasing hormone (GnRH)-stimulated luteinizing hormone (LH) release by anterior pituitary gland and
hCG-stimulated testosterone release by LCs of male rats. These results suggested that catechins stimulated
testosterone production by acting on rat LCs via the mechanism of increasing the action of cAMP, but not P450scc,
StAR protein or the activity of intracellular calcium. EC, one of the catechins increased the testosterone secretion by
rat LCs via the enzyme activities of 17beta-hydroxysteroid dehydrogenase (17beta-HSD).
~0 Citings
457. Isoproterenol and cAMP block ERK phosphorylation and enhance [Ca2+]i increases and oxygen
consumption by muscarinic receptor stimulation in rat parotid and submandibular acinar cells
By Soltoff Stephen P; Hedden Lee
Salivary glands are innervated by sympathetic and parasympathetic neurons, which release neurotransmitters that
promote fluid secretion and exocytosis when they bind to muscarinic and beta-adrenergic receptors, respectively.
Signaling pathways downstream of these receptors are mainly distinct, but there is cross-talk that affects receptor-
dependent events. Here we report that the beta-adrenergic ligand isoproterenol blocks increases in extracellular
signal-related kinase (ERK) phosphorylation, a protein kinase C-dependent event promoted by the muscarinic receptor
ligand carbachol in freshly dispersed rat parotid acinar cells. The inhibitory action of isoproterenol was reproduced by
cAMP stimuli (forskolin) and mimetics (dibutyryl-cAMP, 8-(4-chlorophenylthio)-cAMP), including one highly selective for
protein kinase A (N(6)-benzoyl-cAMP). In contrast, Epac (exchange proteins directly activated by cAMP)-selective
activators did not mimic the blockade of ERK by isoproterenol, suggesting that inhibition involved protein kinase A.
Isoproterenol also blocked ERK downstream of phorbol 12-myristate 13-acetate and the P2X(7) and epidermal growth
factor receptors. Isoproterenol and forskolin blocked MEK phosphorylation, reduced RAF phosphorylation on a
stimulatory site (Ser-338), and increased RAF phosphorylation on an inhibitory site (Ser-259). Inhibitory effects on
ERK were also observed in freshly dispersed rat submandibular acinar cells but not in three immortalized/cancer
salivary cell lines (Par-C10, HSY, HSG), indicating significant differences between native cells and cell lines. Notably,
in native parotid cells isoproterenol enhanced the carbachol-promoted increases in [Ca(2+)](i) and oxygen
consumption, events that initiate and accompany, respectively, the stimulation of fluid secretion by muscarinic ligands.
Thus, isoproterenol produces opposite effects on prominent events downstream of the muscarinic receptor second
messengers diacylglycerol (decrease in ERK phosphorylation) and inositol trisphosphate (increase in [Ca(2+)](i) and
fluid secretion).
~10 Citings
458. Synergistic interactions between sorafenib and bortezomib in hepatocellular carcinoma involve PP2A-
dependent Akt inactivation
By Chen Kuen-Feng; Yu Hui-Chuan; Liu Tsung-Hao; Lee Shoei-Sheng; Chen Pei-Jer; Cheng Ann-Lii
From Journal of hepatology (2010), 52(1), 88-95, Language: English, Database: MEDLINE
BACKGROUND & AIMS: Previously we reported that Akt inactivation determines the sensitivity of hepatocellular
carcinoma (HCC) cells to bortezomib. Here we report that combined treatment with sorafenib and bortezomib shows
synergistic effects in HCC. METHODS: HCC cell lines (PLC/PRF/5, Huh-7, and Hep3B) were treated with sorafenib
and/or bortezomib and analyzed in terms of apoptosis signal transduction. In vivo efficacy was determined in nude
mice with PLC/PRF/5 xenografts. RESULTS: Pretreatment with sorafenib enhanced bortezomib-induced apoptotic
cell death by restoring bortezomib's ability to inactivate Akt in PLC/PRF/5 cells. Knocking down Akt1 by RNA-
interference overcame apoptotic resistance to bortezomib in PLC/PRF/5 cells and ectopic expression of active Akt in
HCC cells abolished the bortezomib sensitizing effect of sorafenib, indicating Akt inactivation plays a key role in
mediating the combinational effects. Moreover, okadaic acid, a protein phosphatase 2A (PP2A) inhibitor, reversed
down-regulation of phospho-Akt (P-Akt) expression induced by co-treatment with sorafenib and bortezomib, and 1, 9
di-deoxy-forskolin, a PP2A agonist, restored bortezomib's effect on P-Akt and apoptosis. Importantly, silencing of
PP2A by RNA-interference reduced the apoptotic effect induced by sorafenib-bortezomib co-treatment, indicating that
PP2A is indispensable for mediating the effects of these drugs. Notably, sorafenib with bortezomib increased PP2A
activity in PLC/PRF/5 cells without altering protein levels of PP2A complex or the interaction between PP2A and Akt
proteins. Finally, sorafenib plus bortezomib significantly suppressed PLC/PRF/5 xenograft tumor growth, down-
regulated P-Akt expression, and up-regulated PP2A activity. CONCLUSIONS: The combination of sorafenib and
bortezomib shows synergy in HCC through PP2A-dependent Akt inactivation.
~12 Citings
459. Defective cAMP generation underlies the sensitivity of CNS neurons to neurofibromatosis-1 heterozygosity
By Brown Jacquelyn A; Gianino Scott M; Gutmann David H
From The Journal of neuroscience : the official journal of the Society for Neuroscience (2010), 30(16), 5579-89,
Individuals with the neurofibromatosis type 1 (NF1) inherited cancer syndrome exhibit neuronal dysfunction that
predominantly affects the CNS. In this report, we demonstrate a unique vulnerability of CNS neurons, but not
peripheral nervous system (PNS) neurons, to reduced Nf1 gene expression. Unlike dorsal root ganglion neurons, Nf1
heterozygous (Nf1+/-) hippocampal and retinal ganglion cell (RGC) neurons have decreased growth cone areas and
neurite lengths, and increased apoptosis compared to their wild-type counterparts. These abnormal Nf1+/- CNS
neuronal phenotypes do not reflect Ras pathway hyperactivation, but rather result from impaired neurofibromin-
mediated cAMP generation. In this regard, elevating cAMP levels with forskolin or rolipram treatment, but not MEK
(MAP kinase kinase) or PI3-K (phosphatidylinositol 3-kinase) inhibition, reverses these abnormalities to wild-type levels
in vitro. In addition, Nf1+/- CNS, but not PNS, neurons exhibit increased apoptosis in response to excitotoxic or
oxidative stress in vitro. Since children with NF1-associated optic gliomas often develop visual loss and Nf1 genetically
engineered mice with optic glioma exhibit RGC neuronal apoptosis in vivo, we further demonstrate that RGC apoptosis
resulting from optic glioma in Nf1 genetically engineered mice is attenuated by rolipram treatment in vivo. Similar to
optic glioma-induced RGC apoptosis, the increased RGC neuronal death in Nf1+/- mice after optic nerve crush injury is
also attenuated by rolipram treatment in vivo. Together, these findings establish a distinctive role for neurofibromin in
CNS neurons with respect to vulnerability to injury, define a CNS-specific neurofibromin intracellular signaling pathway
responsible for neuronal survival, and lay the foundation for future neuroprotective glioma treatment approaches.
~29 Citings
460. Neuropeptide Y Y5 receptor promotes cell growth through extracellular signal-regulated kinase signaling
and cyclic AMP inhibition in a human breast cancer cell line
By Sheriff Sulaiman; Ali Marwan; Yahya Ayesha; Haider Khawaja H; Balasubramaniam Ambikaipakan; Amlal Hassane
Overexpression of neuropeptide Y (NPY) and its receptor system has been reported in various types of cancers. NPY
Y5 receptor (Y5R) has been implicated in cell growth and angiogenesis. However, the role of Y5R in breast cancer is
unknown. To identify the role of Y5R in breast cancer, we screened several breast cancer cell lines to examine the
expression of Y5R and its function in breast cancer. All screened cell lines express both Y1 receptor and Y5R except
BT-549, which expresses mainly Y5R. Binding studies showed that NPY, Y5R-selective agonist peptide, and Y5R-
selective antagonist (CGP71683A) displaced (125)I-PYY binding in BT-549 cell membranes in a dose-dependent
manner. The displacement studies revealed the presence of two binding sites in Y5R with IC(50) values of 29 pmol/L
and 531 nmol/L. NPY inhibited forskolin-stimulated cyclic AMP accumulation with an IC(50) value of 52 pmol/L. NPY
treatment of BT-549 cells induced extracellular signal-regulated kinase phosphorylation but did not alter intracellular
calcium. Y5R activation stimulates BT-549 cell growth, which is inhibited by CGP71683A, pertussis toxin, and
extracellular signal-regulated kinase blockade. CGP71683A alone induced cell death in a time- and dose-dependent
manner in Y5R-expressing cells. The stimulation of MDA MB-231 cell migration by NPY is inhibited by CGP71683A.
Together, our results suggest that Y5R plays an important role in cancer cell growth and migration and could be a
novel therapeutic target for breast cancer.
~7 Citings
461. Kruppel-like factor 4 is widely expressed in the mouse male and female reproductive tract and responds as
an immediate early gene to activation of the protein kinase A in TM4 Sertoli cells
By Godmann M; Kosan C; Behr R
From Reproduction (Cambridge, England) (2010), 139(4), 771-82, Language: English, Database: MEDLINE
Kruppel-like factor 4 (KLF4) is a zinc finger transcription factor critically involved in cell proliferation, differentiation, and
carcinogenesis. Recently, KLF4 has also been used for the generation of induced pluripotent stem cells. In this study,
we analyzed Klf4 expression in different mouse tissues using northern blot analysis and immunohistochemistry.
Focusing on the male and female reproductive tract, we showed for the first time that KLF4 is expressed in the
epithelia of the murine uterus and the vagina. In the male reproductive tract, we detected KLF4 in the epithelia of the
epididymis, ductus deferens, coagulating gland, and the penis. As KLF4 is strongly inducible by FSH signaling in
Sertoli cells and as this transcription factor is also involved in Sertoli cell development, we employed the mouse Sertoli
cell line TM4 as a model system to investigate i) the induction kinetics of Klf4 upon activation of the cAMP/protein
kinase A pathway by forskolin and ii) the effects of Klf4 induction on TM4 cell cycle progression. Interestingly, Klf4
mRNA and protein were rapidly but transiently induced, reaching peak levels after 90-120 min and declining to basal
levels within 4 h. Compared with the inducible cAMP early repressor, an immediate early response gene, the induction
kinetics of Klf4 is much faster. In conclusion, Klf4 is an immediate early gene in TM4 cells and its expression in several
epithelia of the male and female reproductive tract suggests an important role of Klf4 in mouse reproductive functions.
~7 Citings
462. Prostaglandin E2-induced colonic secretion in patients with and without colorectal neoplasia
By Kaltoft Nicolai; Tilotta Maria C; Witte Anne-Barbara; Osbak Philip S; Poulsen Steen S; Bindslev Niels; Hansen Mark
B
From BMC gastroenterology (2010), 109, Language: English, Database: MEDLINE
BACKGROUND: The pathogenesis for colorectal cancer remains unresolved. A growing body of evidence suggests a
direct correlation between cyclooxygenase enzyme expression, prostaglandin E2 metabolism and neoplastic
development. Thus further understanding of the regulation of epithelial functions by prostaglandin E2 is needed. We
hypothesized that patients with colonic neoplasia have altered colonic epithelial ion transport and express functionally
different prostanoid receptor levels in this respect. METHODS: Patients referred for colonoscopy were included and
grouped into patients with and without colorectal neoplasia. Patients without endoscopic findings of neoplasia served
as controls. Biopsy specimens were obtained from normally appearing mucosa in the sigmoid part of colon. Biopsies
were mounted in miniaturized modified Ussing air-suction chambers. Indomethacin (10 microM), various stimulators
and inhibitors of prostanoid receptors and ion transport were subsequently added to the chamber solutions.
Electrogenic ion transport parameters (short circuit current and slope conductance) were recorded. Tissue pathology
and tissue damage before and after experiments was assessed by histology. RESULTS: Baseline short circuit current
and slope conductance did not differ between the two groups. Patients with neoplasia were significantly more sensitive
to indomethacin with a decrease in short circuit current of 15.1 +/- 2.6 microA x cm(-2) compared to controls, who
showed a decrease of 10.5 +/- 2.1 microA x cm(-2) (p = 0.027). Stimulation or inhibition with theophylline, ouabain,
bumetanide, forskolin or the EP receptor agonists prostaglandin E2, butaprost, sulprostone and prostaglandin E1 (OH)
did not differ significantly between the two groups. Histology was with normal findings in both groups.
CONCLUSIONS: Epithelial electrogenic transport is more sensitive to indomethacin in normal colonic mucosa from
patients with previous or present colorectal neoplasia compared to colonic mucosa from control patients. Stimulated
epithelial electrogenic transport through individual prostanoid subtype receptors EP1, EP2, EP3, and EP4 is not
significantly different between neoplasia diseased patients and controls. This indicates that increased indomethacin-
sensitive mechanisms in colonic mucosa from neoplasia diseased patients are not related to differences in functional
expression of EP receptor subtypes.
~1 Citing
463. Protein kinase A and regulation of neonatal Nav1.5 expression in human breast cancer cells: activity-
dependent positive feedback and cellular migration
By Chioni Athina-Myrto; Shao Dongmin; Grose Richard; Djamgoz Mustafa B A
MEDLINE
Voltage-gated Na(+) channels (VGSCs) are expressed in excitable cells (e.g. neurons and muscles), as well as in
some classically 'non-excitable' cells (e.g. fibroblasts), and in carcinomas. In general, functional expression of VGSCs
in plasma membrane (PM) is hierarchical and dynamic. Previously, we have shown that an activity-dependent positive
feedback mechanism involving cAMP-dependent protein kinase A (PKA) plays a significant role in upregulation of
VGSCs in strongly metastatic rat prostate cancer Mat-LyLu cells expressing Nav1.7. Here, we investigated the
possible role of PKA in VGSC regulation and its functional consequences in strongly metastatic human breast cancer
(BCa) MDA-MB-231 cells, where the neonatal splice form of Nav1.5 (nNav1.5) is the predominant VGSC present.
Treatment with the PKA activator forskolin for 24h increased mRNA and PM protein levels of nNav1.5, without
changing the total VGSC protein level. Opposite effects were obtained by application of the PKA inhibitor KT5720 or
the highly specific VGSC blocker tetrodotoxin (TTX), the latter implying activity-dependent upregulation. We tested the
possibility, therefore, that the activity dependence of VGSC (nNav1.5) expression involved PKA. Indeed, TTX
pretreatment reduced the level of phosphorylated PKA and eliminated basal and PKA-stimulated cellular migration.
These data suggested that activity-dependent positive feedback mediated by PKA plays an important role in the
functional expression of nNav1.5 in BCa, and in turn, this enhances the cells' metastatic potential.
~12 Citings
464. Differential modulation of mu-opioid receptor signaling to adenylyl cyclase by regulators of G protein
signaling proteins 4 or 8 and 7 in permeabilised C6 cells is Galpha subtype dependent
By Talbot Jeffery N; Roman David L; Clark Mary J; Roof Rebecca A; Tesmer John J G; Neubig Richard R; Traynor
John R
From Journal of neurochemistry (2010), 112(4), 1026-34, Language: English, Database: MEDLINE
Regulators of G protein signaling (RGS) proteins act as GTPase-accelerating protein to negatively modulate G protein
signaling and are defined by a conserved RGS domain with considerable amino acid diversity. To determine the
effects of specific, purified RGS proteins on mu-opioid signaling, C6 cells stably expressing a mu-opioid receptor were
rendered permeable to proteins by treatment with digitonin. Mu-opioid inhibition of forskolin-stimulated adenylyl
cyclase by [D-Ala(2),N-Me-Phe(4),Gly-ol]-enkephalin (DAMGO), a mu-specific opioid peptide, remained fully intact in
permeabilized cells. Purified RGS domain of RGS4 added to permeabilized cells resulted in a twofold loss in DAMGO
potency but had no effect in cells expressing RGS-insensitive G proteins. The inhibitory effect of DAMGO was
reduced to the same extent by purified RGS4 and RGS8. In contrast, the RGS domain of RGS7 had no effect and
inhibited the action of RGS8 as a result of weak physical association with Galphai2 and minimal GTPase-accelerating
protein activity in C6 cell membranes. These data suggest that differences in conserved RGS domains of specific
RGS proteins contribute to differential regulation of opioid signaling to adenylyl cyclase and that a permeabilized cell
model is useful for studying the effects of specific RGS proteins on aspects of G protein-coupled receptor signaling.
~8 Citings
465. Phosphorylation of VACM-1/Cul5 by protein kinase A regulates its neddylation and antiproliferative effect
By Bradley Shirley E; Johnson Alyssa E; Le Isabelle P; Oosterhouse Elizabeth; Hledin Michael P; Marquez Gabriel A;
Burnatowska-Hledin Maria
Expression of the VACM-1/cul5 gene in endothelial and in cancer cell lines in vitro inhibits cellular proliferation and
decreases phosphorylation of MAPK. Structure-function analysis of the VACM-1 protein sequence identified
consensus sites specific for phosphorylation by protein kinases A and C (PKA and PKC) and a Nedd8 protein
modification site. Mutations at the PKA-specific site in VACM-1/Cul5 ((S730A)VACM-1) sequence resulted in
increased cellular growth and the appearance of a Nedd8-modified VACM-1/Cul5. The aim of this study was to
examine if PKA-dependent phosphorylation of VACM-1/Cul5 controls its neddylation status, phosphorylation by PKC,
and ultimately growth. Our results indicate that in vitro transfection of rat adrenal medullary endothelial cells with anti-
VACM-1-specific small interfering RNA oligonucleotides decreases endogenous VACM-1 protein concentration and
increases cell growth. Western blot analysis of cell lysates immunoprecipitated with an antibody directed against a
PKA-specific phosphorylation site and probed with anti-VACM-1-specific antibody showed that PKA-dependent
phosphorylation of VACM-1 protein was decreased in cells transfected with (S730A)VACM-1 cDNA when compared
with the cytomegalovirus-transfected cells. This change was associated with increased modification of VACM-1
protein by Nedd8. Induction of PKA activity with forskolin reduced modification of VACM-1 protein by Nedd8. Finally,
rat adrenal medullary endothelial cells transfected with (S730A)VACM-1/cul5 cDNA and treated with phorbol 12-
myristate 13-acetate (10 and 100 nm) to induce PKC activity grew significantly faster than the control cells. These
results suggest that the antiproliferative effect of VACM-1/Cul5 is dependent on its posttranslational modifications and
will help in the design of new anticancer therapeutics that target the Nedd8 pathway.
~3 Citings
466. Endoplasmic reticulum stress-induced activation of activating transcription factor 6 decreases cAMP-
stimulated hepatic gluconeogenesis via inhibition of CREB
By Seo Hye-Young; Kim Mi-Kyung; Min Ae-Kyung; Kim Hye-Soon; Ryu Seong-Yeol; Kim Nam-Kyeong; Lee Kyeong
Min; Kim Han-Jong; Choi Hueng-Sik; Lee Ki-Up; et al
The expression of genes encoding key hepatic gluconeogenic enzymes, including phosphoenolpyruvate carboxykinase
(PEPCK) and glucose-6-phosphatase (G6Pase), is regulated at the transcriptional level by a network of transcription
factors and cofactors, including cAMP response element-binding protein (CREB). It has been suggested that
increased endoplasmic reticulum (ER) stress in the liver impairs hepatic glucose metabolism. However, the direct
effect of ER stress on hepatic gluconeogenesis is still not clear. Here, we investigated whether ER stress influences
hepatic gluconeogenesis and whether this process is mediated by activating transcription factor 6 (ATF6) through the
inhibition of cAMP-mediated activation of CREB. A cAMP stimulant, forskolin, and 8-bromoadenosine-cAMP increased
PEPCK and G6Pase mRNA expression in H4IIE rat hepatoma cells, and ER stress induced by tunicamycin or
thapsigargin decreased the expression of these genes in forskolin or 8-bromoadenosine-cAMP-treated cells. In a
transient transfection study, ATF6 inhibited the PEPCK and G6Pase promoters. Also, adenovirus-mediated
overexpression of ATF6 in H4IIE cells decreased forskolin-stimulated PEPCK and G6Pase gene expression.
Moreover, the inhibition of endogenous ATF6 expression by small interfering RNAs restored the ER stress-induced
suppression of PEPCK and G6Pase gene expression. Transient transfection of ATF6 inhibited transactivation by
CREB on the PEPCK and G6Pase promoters, and a gel shift assay showed that Ad-ATF6 inhibits forskolin-stimulated
CREB DNA-binding activity. Finally, we found that expression of ATF6 decreased fasting-induced PEPCK, G6Pase
mRNA expression, and blood glucose levels in mice. Taken together, these data extend our understanding of ER
stress and the regulation of liver gluconeogenesis by ATF6.
~14 Citings
467. Effect of 14-3-3 tau protein on differentiation in BeWo choriocarcinoma cells

By Cheng Y; Hu R; Jin H; Ma K; Zhou S; Cheng H; Ma D; Li X
From Placenta (2010), 31(1), 60-6, Language: English, Database: MEDLINE
This study aimed to investigate the location and function of tau isoform of 14-3-3 proteins in human trophoblast. 14-3-3
tau was localized in human cytotrophoblast cells, but not in syncytiotrophoblast cells in both first trimester and term
placenta by immunochemistry stain. Forskolin-induced cell fusion (BeWo cells) confirmed that 14-3-3 tau was
decreased during trophoblast differentiation. Forskolin-induced differentiation was stimulated by small-interfering (si)
RNA induced down-regulation of 14-3-3 tau, contrarily, it was suppressed by plasmid induced upregulation of 14-3-3
tau in BeWo cells. When BeWo cells were treated with 14-3-3 tau siRNA, an increase in protein concentration of cell
cycle inhibitor p27kip1 and a decrease in protein concentration of proliferating cell nuclear antigen, as well as activation
of the extracellular signal-regulated kinase (ERK) pathway, were also noticed. These findings suggest that 14-3-3 tau
might be mediated trophoblast differentiation through cell cycle regulation.
~1 Citing
468. Expression of mTOR and downstream signalling components in the JEG-3 and BeWo human placental
choriocarcinoma cell lines
By Mparmpakas Dionisis; Zachariades Elena; Foster Helen; Kara Anjeli; Harvey Amanda; Goumenou Anastasia;
Karteris Emmanouil
From International journal of molecular medicine (2010), 25(1), 65-9, Language: English, Database: MEDLINE
Emerging data suggest that nutritional status and body weight are related to reproductive function, and nutrient
imbalances during pregnancy lead to changes in the expression of fetal genes. Recent studies show that the mTOR
acts as a placental growth signalling sensor and its expression is down-regulated in intrauterine growth restriction. To
date, very little is known about the expression of this signalling pathway in choriocarcinoma, one of the most lethal
germ cell cancers. In this study, cultures of fusigenic (BeWo) and non-fusigenic (JEG-3) human choriocarcinoma cell
lines were used to investigate the expression of mTOR and its downstream signalling components. The effects of an
inducer of syncytialisation (forskolin) on mTOR, eIF4E binding proteins (4EBPs) and ribosomal protein S6 kinases
(S6Ks) in BeWo cells were also assessed. RT-PCR studies revealed that mTOR, 4EBP and S6Ks are expressed at
mRNA level in both JEG-3 and BeWo cells. Semi-quantitative RT-PCR analysis revealed that in early stages of
syncytialisation (50 microM forskolin for 48 h), the expression of mTOR and 4EBP was down-regulated when
compared to unstimulated cells. In fully syncytialised cells (50 microM forskolin for 72 h) the expression of both genes
was similar to basal levels. Interestingly, the phosphorylation (Ser371, Thr389) status of p70S6K remained unaltered
upon forskolin treatment. These data validate BeWo cells as an experimental model to study the effects of forskolin-
induced syncytialisation on mTOR signalling.
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