野生型PTEN过表达对体外活化大鼠肝星状细胞ERK信号转导的影响

摘要

Objective The effects of overexpression of wild type phosphatase and tensin homology deleted on chromosome Ten(PTEN)on extracellular signal-regulated kinase (ERK) signal transduction was investigated in activated rat hepatic stellate cell (HSC) in vitro.Methods Using transient transfection technique,adenovirus as a carrier,wild-type PTEN gene was transduced into cultivate activated rat HSC in vitro.Using Western blot and real-time fluorescent quantitation PCR,the expressions of PTEN and ERK1 were detected in HSC.And the expression of phosphorylated ERK (pERK) of HSC was determined using Western blot.Results The exogenous wild type PTEN gene was successfully transduced into activated rat HSC in vitro.The expression of p-ERK was significantly downregulated,P＜0.05,but there were no significant differences in the expression of ERK1 at both transcriptional and translational levels in activated HSC,P＞0.50.Conclusion Through inhibiting the phosphorylation of ERK,the ERK signaling transduction is negatively regulated by the overexpression of wild-type PTEN in activated rat HSC in vitro.%目的 探讨野生型第10号染色体缺失的磷酸酶张力蛋白同源物基因(PTEN)过表达对体外活化大鼠肝星状细胞(HSC)的细胞外信号调节激酶(ERK)信号转导的影响.方法 利用瞬时转染技术,以腺病毒为载体,将野生型P TEN基因转染体外培养的活化大鼠HSC;采用Western blot及实时荧光定量PCR技术检测HSC的PTEN、ERK1蛋白及mRNA表达;并应用Western blot技术检测HSC的磷酸化ERK(p-ERK)表达.结果 外源性野生型PTEN基因在活化HSC大量表达,并显著下调HSC的p-ERK表达(P＜0.05),而对HSC的ERK1蛋白及mRNA表达均无影响(P＞0.50).结论 野生型PTEN过表达通过抑制ERK的磷酸化负性调控体外活化大鼠HSC的ERK信号转导.