Knockdown of fumarate hydratase expression results in apoptosis by inhibition of glucose uptake in HeLa cells

• Main Authors: An-CheihCheng, 鄭安捷
• Other Authors: Wen-Tsan Chang
• Format: Others
• Language: zh-TW
• Published: 2010
• Online Access: http://ndltd.ncl.edu.tw/handle/61032797240498392699

碩士 === 國立成功大學 === 生物化學研究所 === 98 === In 1926, Otto Warburg proposed the theory that certain cancer cells generate ATP dependent on glycolysis pathway rather than oxidative phosphorylation even in the presence of ample oxygen, the phenomenon was called Warburg effect.Mitochondrial defects, hypoxia, oncogenetic signals and altered metabolic enzymes are potential mechanisms leading to increase in glycolysis in cancer cells. Fumarate hydratase (FH) is the component of altered metabolic enzyme. In previous studies, germline mutation of the Kerbs cycle enzyme fumarate hydratase (FH) is related hereditary leiomyomatosis and renal cell cancer (HLRCC).And mutation of FH leads to accumulation of fumarate. The excess metabolite, fumarate inhibits the proline hydroxylases that suppress the expression of hypoxia inducible factor 1 (HIF-1), a transcription factor implicated in tumor angiogenesis and tumor-cell glycolysis. In renal cancer, FH is considered a tumor suppressor gene but rarely studies in other cell line. In this study, I established FH stable knockdown cell lines in HeLa cell and analyzed the differences between wide type cells and stable knockdown cell lines. Suppression of FH has no apparent impact on cell growth, migration ability and mitochondrial membrane potential in normal condition. HeLa-shFH cell lines increase glucose uptake. Then I try to inhibit glycolyis pathway by treatment of 2-deoxyglucose or limit glucose source in HeLa-shFH cell lines and investigate the effect of HeLa-shFH cell lines. I found HeLa-shFH cell lines appear to be more sensitive in low and no glucose condition or in 2-DG existence condition than wild type cell lines. Particularly, in the treatment of 2-deoxyglucose HeLa-shFH cell line prefer apoptosis to necrosis than HeLa cell. HeLa-shFH cell lines and wild type cell lines have the same effect by treatment of complex I inhibitor, Rotenone or cultured in no L-glutamine medium. I demonstrate that HeLa-shFH cell lines depend on glucose.