Effects of PTEN over-expression and its mutant on the apoptosis of activated hepatic steilate cells in vitro
10.3760/cma.j.issn.0254-1432.2009.08.008
- VernacularTitle:PTEN过表达及其突变对体外活化肝星状细胞凋亡的影响
- Author:
Lisen HAO
;
Xiaolan ZHANG
;
Junyan AN
;
Yulin LI
;
Na LIU
;
Huiqing JIANG
;
Xiaopeng TIAN
- Publication Type:Journal Article
- Keywords:
Hepatic stellate cell;
Proliferation;
Apoptosis;
Phosphatase and tensin homology deleted on chromosome ten
- From:
Chinese Journal of Digestion
2009;29(8):529-533
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effects of over-expression of wild-type PTEN gene and its mutant (G129E) on apoptosis and proliferation of activated hepatic stellate cells (HSC) in vitro and its potential mechanisms. Methods The activated HSC cells were cultured in vitro and transfected with PTEN gene and G129E gene via adenoviral vector. The apoptosis of HSC was measured by MTT , and its proliferation was assessed by TUNEL and flow cytometry (FCM) . Western blotting and real-time fluorescent quantitation PCR were used to detect expression of PTEN in HSC. And the changes of Bcl-2 and Bax expression were tested by Western blotting. Results The wild type PTEN gene and G129E gene were successfully transducted into HSC, which resuted in elevated expression of Bax and reduced expression of Bcl-2 (P<0.01). After transduction, HSC proliferation was markedly inhibited with inhibitory rates of 14.03% and 23.12% at 48 and 72 hours in Ad-PTEN ,respectively, as well as 9.52% and 12.63% in Ad-G129E, respectively. Apoptotic rate of HSC exposed to Ad-PTEN or Ad-G129E for 72 hours increased significantly (P<0.01). Furthermore, wild type PTEN was more powerful than G129E for above-mentioned effects. Conclusions Over-expression of wild type PTEN and its mutant G129E can inhibit the proliferation of activated HSC, and induce HSC apoptosis through the Bcl-2/Bax pathway. In addition, the effect of wild type PTEN is more powerful than that of G129E.
