Biokimia Kanker :

Karsinogen, Onkogen & Gen Supresif Tumor

F. Ferdinal

Dept. Biokimia dan Biologi Molekuler

Fakultas Kedokteran
Universitas Tarumanagara
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Pendahuluan
 Kanker dikenal sudah sejak lama sekali !

 Mesir (-2500 SM): osteosarkoma pada mummi.

 Hipocrates (-400 SM) : imbalans antara black

humor dan 3 body humors - natural (alamiah).

 Sir Percival Pott (1775): kanker skrotum.

 Boveri (1914): lesi pada DNA akibat mutasi somatik.

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 Broca (1866): kanker payudara dan kanker hati akibat
kelainan yang sifatnya herediter.
 Peyton Rous (1910): RSV menyebabkan kanker – v-src.
 Bishop & Varmus (1976): v-src punya pasangan homolog
di dalam sel normal – c-src.
 Henry Harris (1969): Gen Supresor Tumor (GST), berperan
dalam mengendalikan pertumbuhan sel.

 Kanker : adalah penyakit genetik (transformasi ganas)

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 Carcinogenesis - Overview

CANCER is an abnormality of cell growth and
multiplication characterised by:


At cellular level
 Excessive cellular proliferation
 Uncoordinated growth
 Tissue infiltration


At molecular level
 Disorder of growth regulatory genes
 Develops in a multistep fashion

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I. Karsinogenesis
A. Agen Penyebab Kanker
1. Faktor Fisika.
Energi Radiasi : sinar-uv, sinar-x, sinar-, sinar kosmis
dapat merusak DNA : langsung atau tidak
langsung.

2. Faktor Kimia.
Senyawa Kimia : sifat-sifat umum :
• struktur • kerusakan DNA
• kerja • mutagen
• metabolisme • inisiasi dan promosi
• ikatan kovalen

3. Faktor Biologis
Virus DNA, Virus RNA, Bakteri & Parasit
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Radiant Energy

Ultraviolet rays, x-rays, and -rays are
mutagenic and carcinogenic


Result in DNA damage
 formation of pyrimidine dimers (elimination of
corresponding bases may form apurinic or
apyrimidinic sites
 single- and double-strand breaks or cross-linking
strands may occur
 x-rays and -rays also cause free radicals to form
in tissues
 resultant OH•, superoxide, and other radicals can
interact with DNA leading to molecular damage and
contribute to carcinogenic effects
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Carcinogens Properties
A. Structure: organic & inorganic
B. Action: Procarcinogen -> Proximate -> Ultimate carcinogen
Electrophiles attact nucleophilic groups
C. Metabolism of chemical carcinogen: CYP450

D. Covalent Binding: to macromolecules, DNA, RNA, protein

Benzo[a]pyrene, 2-Acetylaminofluorene, methyl-4aminoazobenzene
E. Damage to DNA: result in mutation

F. Mutagens: At a molecu­lar level, transitions, transversions.

The Ames assay

G. Initiation and Promotion:Knudson two-hit hypothesis (1971)

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The stages of initiation and promotion of chemical carcinogenesis into skin
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 Tabel 1. Beberapa virus tumor yang penting *
Famili Virus Anggota

 
Virus DNA
Papovavirus Poliomavirus, virus SV40, virus papiloma manusia
(misal: HPV-16)
Adenovirus Adenovirus 12, 18, 31
Herpesvirus Virus Epstein-Barr
Hepadnavirus Virus hepatitis B

Virus RNA
Retrovirus tipe C
Retrovirus tipe B
Virus leukemia dan dan virus sarkoma murin,
Virus leukemia dan sarkoma avian, virus leukema
sel T manusia tipe I dan II
Virus tumor mammae mencit

* Virus penting yang dinyatakan sebagai penyebab tumor pada manusia adalah virus
Epstein-Barr (limfoma Burkitt, kanker nasofaring, limfoma sel B), virus hepatitis B
(karsinoma hepato-seluler), virus papiloma manusia ( berbagai jenis tumor termasuk kanker
cervix) dan virus limfoma-leukemia sel T manusia tipe I (leukemia sel T dewasa).
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B. Perubahan pada transformasi ganas

 Perubahan biokimia
 Perubahan bentuk sel
 Hilangnya inhibisi kontak pertumbuhan
 Hilangnya inhibisi kontak pergerakan
 Hilangnya ketergantungan pada penjangkaran
 Perubahan struktur sitoskeleton
 Berkurangnya kebutuhan faktor pertumbuhan
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II. ONKOGEN
A. Peranan Onkogen Pada Karsinogenesis
 Rous (1910) : Inokulasi virus sarkoma, suatu retrovirus (RSV)
diduga RSV mengandung informasi genetik khusus.

 Vogt & Martin : mutant-delesi RSV replikasi tanpa tranformasi.

RSV mengandung onkogen, sedangkan pada ALV tidak ada.
Onkogen virus : v-src, menyandi protein 60 kD, protein kinase.
 Abelson Leukemia Virus (ALV): Isolasi onkogen Abl. Onkogen
berasal dari sel host.
 Bishop & Varmus (1976) : pelacak cDNA dengan teknik hibridasi.
Membuktikan onkogen v-src berasal dari genom sel host.
 R. Weinberg (1981), transduksi human Ca-bladder pada tikus.
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B. Peran Retrovirus pada Karsinogenesis
Onkogen menyandi berbagai protein yang merupakan komponen
penting dalam lintasan sinyal yang menstimulasi proliferasi sel.

Protein : ~ Faktor Pertumbuhan (GF) : int-1, int-2, sis

~ Reseptor GF : erbB1, fms, kit, met, ret, TrkA
~ Sitoplasmik protein : src, bcr-abl, lck, pim, Ras
~ Protein inti : fos, jun. myc, rel

Contoh : src, suatu tirosin kinase yang bertanggung jawab

atas proses transformasi sel, melalui fosforilasi
abnormal : efek pada vinkulin, glikolisis, lintasan
sinyal PI-3-K dan pompa-Na

Dapat menjelaskan, kenapa virus tumor mempunyai efek pleiotropik

(beragam).

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C. Aktivasi Proto-onkogen menjadi Onkogen
3 mekanisme aktivasi : • Mutasi
• Amplifikasi
• Chromosom Re-arrangement
1. Mutasi
~ Mutasi-delesi, oleh v- onc : erb-B, kit, ros, met, trk  truncated-pp.
~ Mutasi-titik, akibat karsinogen kimia, pada famili (p21) GTP ase.
Contoh, ras : kodon 12 dari GGC (Gly) menjadi GTC (Val)
~ Mutasi-insersi, promoter dan enhancer oleh ALV pada c-myc.
2. Amplifikasi, terjadi melalui replikasi yang tidak sempurna .
~ Contoh pada : myc, erbB-2 dan ras.
3. Chromosomal Re-arrangement, berupa translokasi dan inversi.
~ Aktivasi c-myc pada limfoma Burkitt ( kr. 8  14),
~ Fusi gen :chimerik bcr-abl pada CML ( kr. 9  22 ). Gambar 1,2,3.
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D. Fungsi Produk Onkogen
 Onkogen : GF, GFR, sitoplasmik kinase, protein G dan protein inti.
 Fungsi utama :  regulasi proliferasi sel
 kelangsungan hidup sel (survival cell )
diferensiasi sel
Contoh:

 Kerja faktor pertumbuhan sebagai protein onkogen merupakan

hasil ekspresinya yang abnormal.
 GFR, ujung amino diubah melalui translokasi kromosom, sehingga
terbentuk protein fusi Tel/PDGF
 Onkogen : erb-B, sis, ras, src dan myc.

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E. Gen Supresor Tumor (GST)
GST , menghambat : proliferasi sel & perkembangan tumor.
Berperan sebagai regulasi-negatif.

Henry Harris (1969) : hibrid sel tumor-sel normal, hasil sel normal.
Knudson’s two-hit hypothesis: 2 mutasi pada retinoblastoma herediter Gen Rb
sebagai GST, lokasi pada 13q14, berperan pada siklus sel.
p53, merupakan gen utama dalam menjaga kestabilan genom. fungsi :
aktivator transkripsi, regulasi siklus sel, regulasi apoptosis.
PTEN , pada : melanoma, payudara dan prostat
fungsi: regulasi siklus sel, melalui p27 dan induksi apoptosis.
DPC4, terdapat pada 90% Ca. pankreas, menyandi SMAD, diaktifkan oleh
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 Programmed Cell Death (PCD)

• Proses seluler aktif- fisiologis bagi kematian sel (ACD)

• Fungsi : ~ regulasi proliferasi dan jumlah sel
~ mekanisme proteksi
~ regulasi hubungan antar sel
• Disregulasi : Berbagai akibat.
• Berdasarkan morfologi inti dan biokimia :
1. Apoptosis (klasik) : kondensasi kromatin (+) ; Caspase (+) 2.
Apoptosis-like PCD :kondensasi kromatin -/+ ; Caspase (-) 3. Necrosis-
like PCD : kondensasi kromatin (-) ; Caspase (-) * Tipe lain:
Paraptosis; Dark Cell Death
•Komponen lintasan : pencetus,
modulator / regulator,
efektor
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• Pencetus / stimuli : ekstrinsik dan intrinsik (mitokondria)
Lintasan ekstrinsik: Death Receptor famili TNFR : TNF-R1, Fas, TRAIL-R .
• Regulator utama protein famili Bcl-2 (24 macam)
• pro-apoptosis : Bax, Bak, Bid, Bim, Bmf, Noxa, Puma.
• anti-apoptosis : Bcl-2 dan Bcl-XL
• Efektor : Caspase ( 14 tipe ), inisiator (8,9,10) dan efektor akhir (3,6,7)
calpain, cathepsin

• Komponen Lain:
Protein adaptor: FADD (fas-associated death domain)
TRADD (TNFR-associated death domain)
RIP (receptor interacting protein)
TRAF1(TNFR-associated factor-1) DISC
(Death Inducing Signaling Complex)
• PCD : stimuli, tipe sel dan status metabolik. (F1, F2, F3, F4, F5 )
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Ames Assay
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Activation of Proto-oncogenes


Five mechanisms alter the expression or
structure of proto-oncogenes and participate
in their conversion to oncogenes
 Promoter insertion
 Enhancer insertion
 Chromosomal Translocations Quantitative model
 Gene Amplification
 Point Mutation Qualitative model

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Promoter Insertion

Certain retroviruses lack oncogenes (e.g., avian
leukemia viruses) but may cause cancer over
longer period of time

Infect cell, reverse transcriptase synthesizes
cDNA of RNA genome, integrates into host
genome (provirus)

cDNA of provirus flanked by long-terminal
repeats (LTRs) that can promote transcription

provirus integration near c-myc gene
leads to increased constitutive
expression and formation of B cell
tumor
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Enhancer Insertion

Provirus inserted downstream of myc gene,
or upstream in opposite orientation

myc gene activated because enhancer
sequences in LTR increase rate of
transcription

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 Chromosomal Translocations

Many tumor cells exhibit chromosomal
abnormalities

One type seen in cancer cells is translocation
 piece of one chromosomes is split off and joined to
another chromosome
 if second chromosome donates material to the
first, translocation is “reciprocal”

Burkitt’s lymphoma is fast-growing cancer of
human B lymphocytes
 In certain cases, chromosomes 8 and 14 involved
in reciprocal translocation
 Segment of chromosome 8 that moves to 14
contains c-myc
 Transposition places inactive c-myc under
influence of enhancers from immunoglobulin
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 Gene Amplification


Amplification of certain genes is
found in a number of tumors


Methotrexate, an inhibitor of dihydrofolate reductase
(DHFR), is administered as an anticancer drug
 Tumor cells that become resistant to the action of this
drug amplify the gene for DHFR resulting in increased
enzyme activity
 Appear either as homogeneously staining
regions (HSR) or as self-replicating double-
minute chromosomes (lacking centromeres)

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 Point Mutation


DNA sequence of c-ras proto-
oncogene from normal human cells
and c-ras oncogene from human
bladder cancer showed difference in
only one base (resulting in amino acid
substitution at Gly12)

Mutation results in loss of GTPase
activity, may lead to persistent
activation of MAPK pathway
(mitogenic pathway)

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Representation of a genetic model for colorectal cancer.
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MicroRNA (miRNA) Genes
• Do not encode proteins. Instead, the products of these genes
consist of a single RNA strand of about 21 to 23 nucleotides;
their function is to regulate gene expression.
• Function, depends on their targets in a specific tissue:
- as Oncogene or GST
• Mapping of numerous microRNA genes has shown that many
occur in chromosomal regions that undergo rearrangements,
deletions, and amplifications in cancer cells.
• Expression profiling of microRNA genes associated with:

- tumor classification, diagnosis, staging, and progression,

- as well as prognosis and response to treatment.
• Exampl: - miR-15a and miR-16-1 ....... CLL
- miR21.....x PTEN: breast, lung, prostateJump Ca
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3. Genes involved in DNA repair
• Peroderma pigmentosum,
• Ataxia telangiectasia,
• Fanconi’s anemia and
• Bloom’s syndrome

4. Epigenetic
•Metilasi DNA
•Genomic Imprinting
 The overall progression to malignancy is

therefore a complex event.

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The cell cycle showing checkpoints at which DNA is monitored before the next
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Terima Kasih

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