Analysis of the effect of Aldolase deficiency on cervical cancer cells

• Main Authors: Ying-Chou Chen, 陳盈州
• Other Authors: Wen-Tsan Chang
• Format: Others
• Language: zh-TW
• Published: 2009
• Online Access: http://ndltd.ncl.edu.tw/handle/71622228303319231882

碩士 === 國立成功大學 === 生物化學研究所 === 97 === Otto Warburg addressed a phenomenon that cancer cells prefer the glycolysis pathway to the TCA cycle for obtaining ATP even in aerobic conditions in 1926. Later researches found that some excessive activated oncogenes could mediate the activity of the glycolysis pathway through modulate glycolytic enzymes. Three aldolase isozymes (A, B, and C), encoded by three different genes, are differentially expressed during development. In past suduy, aldolase was known to be upregulated in tumor cells to promote the glycolysis,but reports about aldolase were rare.I used the RNAi-mediated technique to stably knock down the endogenous aldolase A in cervical HeLa cancer model successfully. Analysis of the growth and morphology between wild type and Aldolase A deficient HeLa cells, there does not a difference exist. In immunostaining experiment,although alsdolase is a glycolytic enzyme in the cytoplasma,but I found that Aldolase A also existed in nuclei. Although in normal condition there don’t exist any differences between HeLa-shAldoA and wild cells, we tried give the stress on HeLa-shAldoA cell line. There are some designs: hypoxia and two ETC inhibitors. Because aldolase was induced in hypoxia condition, we treated the cell in hypoxia. In hypoxia experiments, there don’t exist the difference between them. In ETC inhibitors experiments, we found the HeLa-shAldoA was more sensitive than wild cells.We also used two clinical drugs: 2-deoxyglucose and etoposide,the inhibitor of glucose metabolism and topoisomeriase II,respectively.In the treatment of 2-deoxyglucose, shAldoA cells were more sensitive,but not sensitive to etoposide.We speculated that etoposode resulted in the cell death through the genotoxic damage and others are related with the ATP biosynthesis.Three effective drugs,in fact,2-DG inhibits glycolysis and others inhibits ETC,the mechanism involed in cell death of shAldoA was different.In future,we need time to explore what the role aldolase A play on the cell survival.