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目的研究丙型肝炎病毒核心蛋白(HCV-core protein,HCV-C)对HepG2细胞周期、细胞凋亡和p53蛋白表达的影响。方法表达HCV-C的真核表达质粒pcDNA3.1-core,通过Lipofectamine基因转染法转染HepG2细胞,经G418筛选获得稳定转染HepG2细胞,经Western Blot证实HCV核心蛋白阳性表达。利用四甲基偶氮唑蓝比色(MTT)法,平板克隆实验和流式细胞术,蛋白印迹和免疫细胞化学法检测HCV核心蛋白对细胞生长增殖率、细胞周期、细胞凋亡率和p53蛋白表达的影响。结果HCV-C转染组细胞增殖率显著高于空白质粒转染组和未转染组;HCV-C转染组细胞S期所占百分率高于未转染组;HCV-C转染组细胞凋亡率显著低于未转染组;HCV-C转染组细胞突变型p53蛋白的表达高于空白质粒转染组和未转染组。结论HCV核心蛋白可能通过促进突变型p53蛋白的表达,促进HepG2从G0/1期进入S期,促进细胞生长增殖,抑制细胞凋亡。
Objective To investigate the effects of HCV-C core protein on the cell cycle, apoptosis and p53 protein expression in HepG2 cells. Methods The eukaryotic expression plasmid pcDNA3.1-core expressing HCV-C was transfected into HepG2 cells by lipofectamine gene transfection. The stable transfected HepG2 cells were obtained by G418 screening, and the positive expression of HCV core protein was confirmed by Western Blot. The effects of HCV core protein on cell proliferation, cell cycle, apoptosis and p53 were detected by MTT assay, plate clone assay, flow cytometry, Western blotting and immunocytochemistry Effect of protein expression. Results The proliferation rate of HCV-C transfection group was significantly higher than that of blank plasmid transfection group and non-transfection group; the percentage of S phase in HCV-C transfection group was higher than that of untransfected group; The apoptotic rate was significantly lower than that of untransfected cells. The expression of mutant p53 protein in HCV-C transfected cells was higher than that of blank transfected cells and untransfected cells. Conclusion HCV core protein may promote the expression of mutant p53 protein and promote the entry of HepG2 into S phase from G0 / 1 phase, promote cell proliferation and inhibit apoptosis.