Stephanie Zeibak

Advanced Biochemistry
Steiger
09.27.16
Different Mechanisms of Regulation of the Warburg Effect in Lymphoblastoid and Burkitt Lymphoma Cells
Burkitt lymphoma (BL), a B-cell derived childhood malignancy, is known to be associated with
Epstein- Barr virus (EBV). Along with the virus is an increase in the production of MYC protein, which is
involved in many different pathways, including that of glycolysis. Glucose metabolism is known to be
different in normal cells vs tumor cells. Normal cells convert one glucose molecule to two pyruvate
molecules, which then in turn produces ATP. In tumor/ abnormal cells, the pyruvate is converted to lactate
through LDHA- a process usually done due to low oxygen- while oxygen is still present, causing the
Warburg Effect. Scientists have discovered that the regulation of this aerobic glycolysis in LCLs is due
to HIF1A; however, many genes are targeted by MYC, not HIF1A. The main questions in the study is
whether MYC or HIF1A is the main regulator of expression in aerobic glycolysis in BL lines is and if it is
the cause of the Warburg effect in BL cells.

A variety of methods- western blot, quantitative PCR, cell culture, statistical analysis, and
biochemical assays- were used to answer these questions. The western blot was performed to assess the
expression of HIF1A at a protein level. The use of spectrophotometry showed that pyruvate
concentrations were higher in BL lines than LCLs suggesting that BLs proliferate faster than LCLs. After
conducting a PCR, it was established that HIF1A had little effect on BLs compared to its effect on LCLs,
as well as the finding that MYC functions as a transcription factor in BLs, but not in LCLs. Further,
LDHA activity was assessed for BLs and LCLs to find that aerobic glycolysis was controlled by MYC in
BL cells. The study finds that the HIF1A protein was expressed at high levels in EBV- positive BLs. To
understand the importance of HIF1A on BLs, the protein was affected by ACF. This resulted in LCLs
being affected, whereas BLs showed no inhibition. This allowed the author to conclude that HIF1A is not

a major regulator in BLs. MYC, on the other hand, is another transcription factor involved. To observe the
effects it has, it was treated with 10058-F4. The results showed no change in LCL levels; however, BLs
showed a decreased LDHA.

These results can allow the author to conclude that HIF1A is not a major regulator in BL cells, but
is for LCLs. Another conclusion that can be made is that high levels of MYC can be attributed to the
Warburg effect in BLs and high levels of HIF1A can be attributed to the Warburg effect in LCLs. This is
what interested me about the study. At first, it was a bit difficult to understand the different techniques use
and why, but after reading it multiple times, it was interesting to see just how to different proteins could
affect two different cell lines to create the same Warburg Effect. No questions were left unanswered by
the scientists; however, I am left with some questions. Is MYC and HIF1A the only two that could affect
BLs and LCLs?

Reference
https://www.cellsignal.com/contents/science-pathway-research-cellular-metabolism/warburg-effectsignaling-pathway/pathways-warburg
Mushtaq M, Darekar S, Klein G, Kashuba E (2015) Different Mechanisms of Regulation of the Warburg
Effect in Lymphoblastoid and Burkitt Lymphoma Cells. PLoS ONE 10(8): e013614