PS1TP2 On The Influence Of HepG2 Cells Proliferation And Apoptosis And Mechanism Research

Objective: To investigate the effects of Pre-S1 protein transactivator gene 2(PS1TP2)on proliferation and apoptosis of hepatocellular carcinoma HepG2 cells,and to explore the role of PS1TP2 in the development and progression of hepatocarcinoma associated with HBV infection and its possible mechanism.Methods: First,the basic expression levels of PS1TP2 gene in normal liver cell line L02 cells and hepatoma cell line HepG2 cells were detected by real-time quantitative PCR(RT-PCR).After chemically synthesizing the three batches of small interference RNA(si PS1TP2)of the PS1TP2 gene and the universal negative control(siNC)with no homology to the target sequence,RT-PCR was used to screen the si PS1TP2 with the best interference effect.For the subsequent experiments;siPS1TP2 and its negative control siNC were transiently transfected into HepG2 cells,divided into interference group(siPS1TP2)and control group(siNC).After 48 hours of routine culture,the following experiment was performed.1.RT-PCR and Western Blot were used to verify the successful interference of PS1TP2 gene expression in HepG2 cells.2.The cell proliferation activity detection kit(CCK8 kit)was used to detect the proliferation activity of HepG2 cells in the two groups.3.Using AnnexinV/7AAD Flow cytometry was used to analyze the cell count after cell staining,and the number of apoptosis between the two groups was detected.4.The expression levels of Bcl-2 and Bax mRNA were detected by RT-PCR;5.Western The expression levels of Bcl-2,Bax,AMPK and m-TOR protein were detected by Blot method,and the ratio of Bcl-2/Bax was calculated.6.The activity of caspase-3 was detected by chemiluminescence method.Results:1.RT-PCR confirmed that the mRNA expression level of PS1TP2 gene in HepG2 cells was significantly higher than that in normal liver cell line L02;2.RT-PCR and Western blot confirmed that the expression of PS1TP2 mRNA and protein was successfully interfered(P<0.05);3.Compared with the siNC control group,the proliferation activity of CCK-8 was significantly decreased in HepG2 cells that interfered with PS1TP2 expression(P<0.05).The number of Annexin V+ cells was significantly increased in HepG2 cells interfering with PS1TP2 expression.(P<0.01);4.Compared with the siNC control group,the expression of Bcl-2 mRNA in HepG2 cells was significantly decreased(P<0.05),and the expression of Bax mRNA was significantly increased(P<0.05).Compared with the siNC control group,the expression of Bcl-2 and m-TOR protein in HepG2 cells was significantly decreased(P<0.01),and the expression of Bax and AMPK protein was significantly increased(P<0.05).The ratio of Bcl-2/Bax decreased significantly(P<0.05).6.Compared with the siNC control group,the activity of Caspase-3 was significantly increased in HepG2 cells expressing PS1TP2(P<0.05).Conclusion: 1.Interference with PS1TP2 expression can promote apoptosis of HepG2 cells via mitochondrial pathway,and its effect on apoptosis may be achieved by down-regulating the ratio of Bcl-2/Bax.2.Interference with PS1TP2 expression may inhibit the proliferation of HepG2 cells by regulating the AMPK-m-TOR pathway.3.PS1TP2 may participate in the occurrence and development of liver cancer caused by HBV infection by affecting the balance of apoptosis and proliferation.