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目的探讨丙型肝炎病毒非结构蛋白质5A(HCV NS5A)对Survivin基因表达的影响。方法用含HCV NS5A基因的表达质粒(pCNS5A)转染HepG2细胞,采用免疫细胞化学技术检测HCV NS5A蛋白质的表达,采用逆转录聚合酶链反应和Western blot方法观察HCV NS5A对Survivin mRNA和蛋白质表达水平的影响。结果转染pCNS5A 的HepG2细胞胞浆可见HCV NS5A蛋白质的表达。转染pCNS5A的HepG2细胞内Survivin mRNA和蛋白质表达均比转染空白载体(pRc/CMV)的HepG2细胞及未转染质粒的HepG2细胞明显增强,而后两者Survivin mRNA和蛋白质表达相似。结论HCV NS5A可激活Survivin蛋白质表达,其途径可能通过调节Survivin基因的转录水平(mRNA转录增强)来实现。
Objective To investigate the effect of hepatitis C virus nonstructural protein 5A (HCV NS5A) on Survivin gene expression. Methods HepG2 cells were transfected with the expression vector pCNS5A containing HCV NS5A gene and the expression of HCV NS5A protein was detected by immunocytochemistry. The expression of Survivin mRNA and protein was detected by reverse transcriptase-polymerase chain reaction and Western blot. Impact. Results The expression of HCV NS5A protein was observed in the cytoplasm of HepG2 cells transfected with pCNS5A. The expression of Survivin mRNA and protein in HepG2 cells transfected with pCNS5A was significantly higher than that in HepG2 cells transfected with blank vector (pRc / CMV) and untransfected plasmids, while the expression of Survivin mRNA and protein in the latter two were similar. Conclusion HCV NS5A can activate Survivin protein expression, which may be through regulating the transcription level of Survivin gene (mRNA transcription enhancement).