Effect Of RNA Silencing CtBP1on The Biological Behavior Of Human Hepatoma Cell Line HepG2

Objective:To explore the effects of small interference RNA (siRNA) on the C-terminal-binding protein1(CtBP1) and its target gene E-cadherin of human Hepatoma Cell Line HepG2. To explore the effect of siRNA on the cell proliferation, cell cycle, cell apoptosis and invasiveness of HepG2. Discuss the role of CtBPl in the occurrence and development of liver cancer. Provide the theoratical basis for the further research about the molecular mechanism and gene therapy.Methods:the cultured human HepG2cells were transfected with control siRNA-A by X-treme GENE siRNA Transfection Reagent. Explore the transfection rate by inverted fluorescence microscope. The HepG2cells were divided into blank control group, transfection reagent group(X-treme GENE siRNA reagent alone), control siRNA-A group(negative control) and interference group, and were transfected with control siRNA-A and CtBP1siRNA(h) by X-treme GENE siRNA Transfection Reagent. The expression of CtBP1mRNA and E-cadherin mRNA and their related protein were detected by RT-PCR and western blotting. The cell proliferation of HepG2was detected by MTT. The cell cycle and apoptosis were detected by flow cytometry. The invasiveness of HepG2was detected by transwell chamber.Result:HepG2was transfected by siRNA successfully with a transfection rate of85%. The HepG2cells in the interference group revealed down-regulated expression of CtBP1mRNA and CtBP1(p<0.05). The inhibition rate of CtBP1mRNA and its protein were38.11%and62.39%separately, while the difference among other groups have no statistics significant(p>0.05). The HepG2cells in the interference group revealed up-regulated expression of E-cadherin mRNA and its protein(p<0.05) while the other groups show no statistics significant(p>0.05). The cell proliferation of HepG2in the interference group was inhibited significantly. The inhibition rate were22.34%(24h),52.15%(48h),53.97%(72h),54.77%(96h) after the transfect, as compared with the other three groups(p<0.05). the cell numeric of every period of the cell cycle shows no different among4groups (p>0.05). The hepG2cells in the interference group has a high cell apoptosis rate. The invasiveness of hepG2cells in the interference group was also obviously decreased(p<0.05).Conclusion:HepG2was transfected by CtBPl siRNA successfully. CtBPl siRNA(h) can inhibit the expression of the CtBP1gene of Human Hepatoma Cell Line HepG2and up-regulate the expression of E-cadherin. CtBP1siRNA(h) can inhibit the proliferation, promote the apoptosis, decrease the invasiveness of HepG2, while has no effect on the cell cycle. CtBP1plays an important role in the occurrence and development, invasion and metastasis of tumor. CtBP1may be a potential new target gene for anticancer therapy.