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背景与目的:探讨乙型肝炎病毒X蛋白在细胞凋亡中的作用。材料与方法:采用基因重组技术构建乙型肝炎病毒x基因的真核表达载体pcDNA3-HBx。脂质体FuGE NE6转染HeLa细胞,并用蛋白印迹检测pcDNA3-HBx在HeLa细胞中的表达。流式细胞术检测细胞凋亡率。结果:成功构建了可在真核细胞中稳定表达X蛋白的HBx基因真核表达载体pcDNA3-HBx。稳定转染pcDNA3-HBx DNA的HeLa-Xs细胞的凋亡率为9.8%,较空白HeLa细胞的凋亡率(0.4%)以及对照细胞HeLa-Vs细胞的凋亡率(1.3%)高;两只转基因小鼠肝细胞的细胞凋亡率分别为54.4%和40.4%,较正常小鼠肝细胞的细胞凋亡率(6.8%)明显增高。结论:X蛋白可在体外和体内诱导细胞凋亡。
BACKGROUND AND AIM: To investigate the role of hepatitis B virus X protein in apoptosis. Materials and Methods: The eukaryotic expression vector pcDNA3-HBx of hepatitis B virus x gene was constructed by gene recombination technology. The liposome FuGE NE6 was transfected into HeLa cells and Western blot was used to detect the expression of pcDNA3-HBx in HeLa cells. Flow cytometry was used to detect the apoptosis rate. Results: The HBx gene eukaryotic expression vector pcDNA3-HBx, which can stably express X protein in eukaryotic cells, was successfully constructed. The apoptosis rate of HeLa-Xs cells stably transfected with pcDNA3-HBx DNA was 9.8%, which was higher than that of blank HeLa cells (0.4%) and the apoptosis rate of control cells HeLa-Vs cells (1.3%); The apoptotic rate of hepatocytes in transgenic mice was 54.4% and 40.4%, respectively, which was significantly higher than that in normal mice (6.8%). CONCLUSION: X protein induces apoptosis in vitro and in vivo.