Effect of HSG gene on proliferation and apoptosis of rat hepatic stellate cell line HSC-T6

Objective To discuss the effects of HSG gene on proliferation and apoptosis of rat hepatic stellate cell line HSC-T6.Methods To construct the recombinant retroviral vector carrying rat HSG gene,transient transfection into rat hepatic stellate cell line HSC-T6,real-time quantitative polymerase chain reaction (RT-PCR) and Western blotting were used to detect the mRNA expression and protein levels of HSG respectively.Cell counting kit-8 (CCK-8) assay was used to investigate cell proliferation activity.Flow cytometry was used to measure the cell cycle progression and cell apoptosis.Effect of HSG on HSC-T6 cells proliferation were investigated by Western blotting for the expression of cyclin dependent kinase inhibitors p21,p27 and proliferating cell nuclear antigen (PCNA).Results Compared with blank control group and negative control group,the protein and mRNA expression (14.93 ± 1.10 vs.1.03 ± 0.33,14.93 ± 1.10 vs.1.00 ± 0.15) of HSG was significantly increased in transfection group (all P ＜ 0.05).Meahwhile,the cell proliferation activity remarkably decreased in transfection group (both P ＜ 0.05).Furthermore,cell cycle arrested in the G0/G1 phase with the increasing apoptosis rate [(23.88 ± 0.96) ％ vs.(3.38±0.16)％,(23.88±0.96)％ vs.(3.54 ±0.24)％] as well as the expression of p21 and p27,and the reducing expression of PCNA (all P ＜ 0.05).Conclusion Overexpression of HSG could inhibit the proliferation of rat hepatic stellate cell line HSC-T6,the cell cycle arrest in the G0/G1 phase and induce its apoptosis.HSG may play a role by participating in the regulation of cell cycle. Key words: Hepatic fibrosis ;  HSG ;  Transient transfection ;  Cell cycle ;  Cell apoptosis