Specific Mcl-1 SiRNA Inhibits Mcl-1 Expression In HCC HepG2 Cells And The Anti-apoptosis Function Of Mcl-1 In Vitro

AIM:The current studies were designed to investigate the effects of specific mcl-1 siRNA on mcl-1 expression in HCC HepG2 and effects of specific mcl-1 siRNA on proliferation and apoptosis of HCC HepG2 in vitro.Discuss the role of mcl-1 on apoptosis of HepG2.METHODS:1 Through counting method to set out growth curve of HepG2 cells,and understand their basic characteristics.2 Human HCC cell line HepG2 was cultured in vitro and transfected with two siRNAs(different loci) of mcl-1 by Lipofect AMINE 2000. The level of mcl-1 mRNA expression before and after transfecting was detected by semi-quantitative reverse transcription polymerase chain reaction(RT-PCR).The interfering effect between the two siRNAs and the specificity and time effect for interference were compared.Then the level of Mcl-1 protein expression before and after interfering was detected by Western blot.3 Through counting method to set out growth curve of HepG2 before and after transfecting. Apoptosis of HepG2 cells before and after transfecting was determined by Annexin V/PI.RESULTS:1 Through counting method,growth curve of HepG2 was set out.2 The level of mcl-1 mRNA in HepG2 was inhibited by the specific siRNAs.The decrease of mcl-1 mRNA expression began to appear 24 h after transfection.And the most apparent interfering efficiency was (67.25±4.47)% 48 h after transfection,which was markedly higher than that in the cells transfected with the control siRNA (P<0.05).Both siRNAs(siRNA-F1 and siRNA-F2) had interfering effect on mcl-1 mRNA expression,but there was only a little diference between them.Western blot showed that expression of Mcl-1 protein was significantly downregulated after interfering with siRNA-F1(P<0.05).3 Proliferation of HepG2 was inhibited after transfecting with specific mcl-1 siRNA-F1. Apoptotic rate was significantly increased after transfecting with specific mcl-1 siRNA-F1 determined by Annexin V/PI(P<0.01).CONCLUSION:1 Specific mcl-1 siRNA can downregulate the level of mcl-1 mRNA in HepG2 cells.And Mcl-1 protein expression is also downregulated by specific mcl-1 siRNA.2 Proliferation of HepG2 cells was inhibited after transfecting with specific mcl-1 siRNA-F1.Apoptotic rate was significantly higher in cells transfected with siRNA-F1 comparing to other cells,which indicated that Mcl-1 may play a role of anti-apotosis in HepG2 cells in vitro.