Correlation Between HSP90, SIRT3 Gene Polymorphism And The Risk Of Liver Cancer And Between Their Expression And Invasion In Hepatoma Cells

Object By studying the expression of HSP90 and SIRT3 in hepatocellular carcinoma and their correlation between the polypeptide of the site and the risk of hepatocellular carcinoma, we try to discover and identify of new susceptible genes in hepatocellular carcinoma. To provide new markers for the vulnerable populations of liver cancer prevention, and to provied new tragets for the development of hepatocellular carcinoma and clinical development of new drugs. so as to improve the prevention and treatment effect of HCC.Methods(1)We selected 198 cases of liver cancer patients in this hospital in January 2013-October 2015 period, between the ages of 35-76 years old. We also collected 198 cases of healthy volunteers as controls from medical centers, between the ages of 33-69 years old. All subjects gave written informed consent. We analyzed the correlation between HSP90, SIRT3 gene polymorphism and the risk of liver cancer, in addition to analyzing the correlation between the drinking history of patients with liver cancer susceptibility.(2)We use p GCSIL-GFP-HSP90 recombinant lentiviral vector was transfected into human hepatoma AMMC-7721 cells, HSP90 m RNA expression was detected in cells after transfection by fluorescence quantitative PCR; MTT assay cell growth; Apoptosis is detected by flow cytometry; invasive ability of the cells to be detected by Transwell chamber method; proliferative capacity of the cells were detected by colony formation test; migration of cells was detected by cell scratch assay; apoptosis was detected by Caspase- 3 activity detection experiments.(3)Expression of HSP90 gene is silenced in AMMC-7721 cells were transfected with RNA interference, to build HSP90 gene silencing cell line si HSP90-AMMC-7721.WT-HSP90 wild-type and mutant Mut-HSP90 was transfected into si HSP90-AMMC-7721 cells; expression of SIRT3 m RNA in cells was detected by fluorescence quantitative PCR; expression of SIRT3 protein in the cells was detected by western blot method.Results(1)There were certain positive correlations between HSP90 rs8005905、 rs3742429 polymorphism and the risk of liver cancer, There were alslo significant correlation between HSP90 rs3742429 liver cancer who with a history of drinking.(2)HSP90 expression increased in AMMC-7721 cells, can significantly improve the ability of proliferation; flow cytometry results showed that the expression of HSP90 increased apoptosis in cells not AMMC-7721 cells; Transwell chamber test results show that the method, increased HSP90 expression can significantly increase the number of invasive cells; colony formation assay results indicated that increased in AMMC-7721 cells, the ability to form cell clones significantly increased HSP90 expression; the expression of cell scratch test results show that increased HSP90 can significantly improved migration AMMC-7721 cells; caspase-3 activity assay results indicate that increased expression of HSP90 in AMMC-7721 cells does not affect caspase-3 activity.(3)Expression of SIRT3 protein was detected by fluorescence quantitative PCR and western blot, results showed that the expression of SIRT3 m RNA and the corresponding protein were significantly decreased by transfected with wild-type HSP90 plasmid in si HSP90-AMMC-772 cells; SIRT3 m RNA and the corresponding protein had on significant differences after transfection with mutant HSP90 in si HSP90-AMMC-7721 cells.Conclusion HSP90 gene polymorphism and the patients with a history of alcohol were related to liver cancer, and HSP90 could promote the proliferation, invasion and metastasis of liver cancer cells, this process was by suppressing the expression of SIRT3 implemented.