HCV NS5A反式激活基因NS5ATP13对肝癌细胞生长的影响

Growth-promoting effects of NS5ATP13 transactivated by HCV NS5A on hepatoma cells

目的验证HCV NS5A对NS5ATP13基因反式激活作用,并检测NS5ATP13基因表达产物对Huh-7细胞系生长的影响。方法应用实时定量逆转录聚合酶链反应(RT-PCR)方法验证HCV NS5A对NS5ATP13基因的反式激活作用,构建NS5ATP13与绿色荧光蛋白(GFP)的融合蛋白表达载体,用荧光显微镜观察NS5ATP13-GFP融合蛋白在细胞的分布。结果 HCV NS5A可反式激活NS5ATP13基因的表达,NS5ATP13-GFP融合蛋白荧光主要集中在细胞核质,转染细胞出现大量双核及多核现象,活细胞发光法检测NS5ATP13具有促进细胞增殖的作用。结论 NS5ATP13基因生物学功能的研究,为HCV致肝纤维化和肝癌的病理机制研究提供新的思路。

Objective To testify the transactivated role of HCV NS5A to NS5ATP13 gene and the effect of NS5ATP13 gene expression on Huh-7 cells. Methods RT-PCR was carried out to testify the transactivated role of HCV NS5A to NS5ATP13 gene and NS5ATP13-GFP fusion gene expression vector was constructed. The subcellular localization of NS5ATP13-GFP fusion protein was observed by fluorescence microscope. Results HCV NS5A can transactivate the NS5ATP13 gene expression, NS5ATP13-GFP fusion protein mainly located in the nucleus and most of the cell occured two or more nucleus. Celltiter-Glo luminescent cell viability assay showed that the growth of Huh-7 cells transfected with NS5ATP13 gene was dramatically promoted compared with the parent Huh-7 cells. Conclusions The biological functional study of NS5ATP13 brought some new clues for studying the pathogenesis of HCV induced hepatic fi brosis and hepatoma.