[The mechanisms of inhibitory effect of adenovirus-mediated wild-type PTEN gene on the proliferation in activated hepatic stellate cells in vitro].

Objective Using an adenoviral vector,the wild-type PTEN gene was transduced into activated hepatic stellate cell (HSC) culcured in vitro and cell cycle markers and were detect.Thereby,the potential mechanisms of inhibitory effect of the wild-type PTEN overexpression on the proliferation in activated HSC was investigated.Methods The wild type PTEN gene was transduced into activated HSC (HSC-T6) cultured in vitro mediated by adenoviral vector.PTEN expression in HSC was measured by Western blot and Real-time fluorescent quantitation PCP.Flow cytometry (FCM) was then used to detect cell cycle phase of activated HSC.And the expressions of cyclinD1 and cyclin dependent kinase 4 (CDK4)in HSC were determined by Western blot.Results The data showed that exogenous wild type PTEN gene was successfully transduced and expressed in activated HSC cultured in vitro.The over-expression of wild type PTEN resulted in the increased number of HSC at G0/G1 phase (P＜ 0.01),and the number of HSC at S phase and G2/M phase were decreased significantly,P＜0.01.Furthermore,there were decreased cyclinD1 and CDK4 expression in HSC infected with Ad-PTEN,P＜ 0.01.Conclusion The over-expression of wild type PTEN inhibit transition of activated HSC in vitro from G1 to S phase and arrest cell cycle of them at G0/G1 phase via the down-regulated expressions of cyclinD 1 and CDK4,and then inhibit HSC proliferation. Key words: Liver cirrhosis;  Cell proliferation;  Cell cycle;  Hepatic stellate cell;  PTEN