乙型肝炎病毒X蛋白表达及其对HepG2细胞凋亡的影响

Effects of expression of HBV X gene on apoptosis of HepG2

目的探讨HBVX蛋白表达对转染x基因HepG2细胞的影响。方法构建真核表达质粒pCD-NA3.1(+)/x,磷酸钙沉淀法转染到人肝癌细胞株HepG2;Western-blot检测X蛋白表达;流式细胞仪检测细胞周期和凋亡率;MTT法测定细胞活力。结果磷酸钙沉淀法能有效的导入x基因且Western-blot能检测到X蛋白表达;MTT法、流式细胞术结果显示X蛋白表达组细胞活力受抑制(P≤0.01),凋亡率增加(P≤0.01)。结论HBV感染及其相关慢性肝病中,X蛋白的表达可能与肝细胞凋亡,炎症活性相关。

[Objective] To elucidate the effect of the expression of HBV X gene on hepatocyte apoptosis. [Methods] The eukaryotic expression vector of x gene（pCDNA3.1-x） was constructed and introduced into human hepatocarcinoma cell strain HepG2 with calcium phosphate precipitates transfection. MTT colorimetric assay and flow cytometric analysis of DNA histograms were used to measure the proliferation and cell cycle distribution and cell apoptosis of HepG2 cells. Western-blot was used to confirm the expression of recombinant X protein. [Results] The expression of HBV X protein exerted the inhibitory actions on HepG2 cells（P≤0.01）. The increasing apoptotic rate of HepG2 was significant compared with the controls （P≤0.01）. [Conclusion] HBV x gene and X protein can promote the apoptosis in HepG2 cells.