目的 研究丙肝病毒(HCV)蛋白NSSA对PI3K/Akt信号的调节机制及其意义.方法 HepG2细胞分别转染NS5A质粒和对照载体.提取总蛋白,用Western blotting法分析PI3K信号Akt磷酸化水平,并用免疫沉淀法检测p85酪氨酸磷酸化水平及p85与p110蛋白间的相互作用.结果 NS5A转染细胞pAkt蛋白水平上调,同时p85酪氨酸磷酸化水平显著提高,但催化亚基p110与调节亚基p85的结合作用没有明显变化.结论 丙肝病毒(HCV)蛋白NSSA可以和PI3K p85亚基结合而调节PI3K/Akt信号通路,但其机制可能有p85/p110以外的机制.这可能为临床丙肝IFN敏感性的诊断与治疗提供依据.%Objective To investigate the molecular mechanism of NS5A regulation of PI3K/Akt signaling. Methods HepG2 cells were transfected with NS5A(wt) plasmids or control plastnids. Akt phosphorylation was tested in cell lysates using Western blotting. P85 tyrosine phosphorylation and the interaction between p85 and p110 did not change. Results Overexpression of NS5A increased Akt phosphorylation. Consistently the p85 tyrosine phosphorylation was enhanced. However, no significant change of interaction between pS5 and p110 was observed. Conclusion NS5A regulation of PI3K/Akt may have more mechanism than via p110 , which may have implication for HCV responses to IFN and the clinical diagnosis.
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