摘要
目的:研究HCV-NS4B对肝细胞Caspase-3及Survivin表达的影响,探讨HCV-NS4B对肝细胞凋亡的影响及相关机制。方法:通过转染丙型肝炎病毒非结构蛋白4B(HCV-NS4B)至LO2肝细胞中,设置三组细胞(空白对照组、空白载体PCXN2组、PCXN2-NS4B组),利用流式细胞仪检测各组Caspase-3的表达情况,免疫组化方法观察三组中Survivin的表达情况。结果:PCXN2-NS4B转染入LO2细胞并有稳定表达,流式细胞术检测各组Caspase-3的表达情况:空白对照组、转染PCXN2组及PCXN2-NS4B组均有Caspase-3表达,分别为:(23.45±1.57)%、(23.17±1.79)%及(4.34±0.59)%,空白对照组、转染PCXN2两组差异无统计学意义(P>0.05),而空白对照组、转染PCXN2组与PCXN2-NS4B组间比较差异有统计学意义(P<0.01)。免疫组化方法检测Survivin的表达情况:空白对照组阳性表达率为25%,PCXN2组表达率为16.67%,PCXN2-NS4B组表达率为75%,空白对照组、转染PCXN2两组差异无统计学意义(P>0.05),而空白对照组、转染PCXN2组与PCXN2-NS4B组间比较差异有统计学意义(P<0.05)。结论:①NS4B转染入肝细胞后有稳定表达;②NS4B可抑制Caspase-3表达、促进Sur-vivin的表达;③HCV-NS4B可能抑制Caspase-3表达、促进Survivin的表达而抑制肝细胞凋亡。
Objective：To investigate the effect of hepatitis C virus non-structural protein 4B （HCV-NS4B） on Survivin,and Caspase-3 expression,and to explore its effect on Apoptosis and proliferation of hepatoeyte and its possible mechanism. Methods： By transfecting the recombinant plasmid （PCXN2-NS4B） into LO2 liver cells with liposome, the experiment was divided into ： LO2 liver cells group, empty vector PCXN2 group and PCXN2-NS4B group. Flow cytometry was used to determine the expression of Caspase-3,and immunohistochemical method was used to detect the expression of Survivin. ResuIts：PCXN2-NS4B was transfected into LO2 ceils, with a stable expression.Caspase-3 expression ： in LO2 liver cell group, empty vector PCXN2 group and PCXN2-NS4B group was （23.45 ±1.57）%,（23.17 ± 1.79）% and （4.34 ± 0.59）% respectively.The difference between LO2 liver cell group and empty vector PCXN2 group was not significant （P〉0.05）, while between LO2 liver cell group, and PCXN2-NS4B group there was significant difference （P〈0.01）.Positive rate of Survivin expression in LO2 liver cells group, PCXN2 group and PCXN2-NS4B group was 25%,16.67% and 75% respectively. There was no significant difference between LO2 liver cells group and empty vector PCXN2 group （P〉0.05）; but between LO2 liver cells group and PCXN2-NS4B group there was significant difference （P〈0.05）.Conelusions： ①Stable expression of the recombinant plasmid is found in transfected L02 liver ceils.②HCV-NS4B inhibits Caspase-3 expression and promotes the expression of Survivin. ③HCV-NS4B Inhibits liver cell apoptosis via inhibiting expression of Caspase-3 and increasing expression of Survivin.
出处
《泸州医学院学报》
2009年第4期339-343,共5页
Journal of Luzhou Medical College
基金
四川省教育厅资助课题(07ZA148)
