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目的:初步探讨HBV X蛋白表达对转染X基因HepG2细胞的影响。方法:磷酸钙沉淀法转染真核表达质粒pCDNA3.1(+)/x到人肝癌细胞株HepG2;72 h后,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 investigate the effect of HBV X protein expression on HepG2 cells transfected with X gene. METHODS: Western Blot was used to detect the expression of X protein after transfection with eukaryotic expression plasmid pCDNA3.1 (+) / x to human hepatocellular carcinoma cell line HepG2 by calcium phosphate precipitation method. The cell cycle and apoptosis rate were detected by flow cytometry MTT assay was used to determine cell viability. Results: The calcium phosphate precipitation method could effectively introduce X gene and the expression of X protein was detected by Western-blot. MTT assay and flow cytometry showed that the cell viability was inhibited (P <0.01) and the apoptosis rate was increased (P <0.01). Conclusion: The expression of X protein in HBV infection and its related chronic liver diseases may be related to hepatocyte apoptosis and inflammatory activity.