乙肝病毒X蛋白诱导HeLa细胞和转基因小鼠肝细胞凋亡的研究

Apoptosis Induced by Hepatitis B Virus X Protein in HeLa Cells and Transgenic Mice Hepatocytes

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 ＆ AIM： To study the role of hepatitis B virus X protein in apoptosis. MATERIALS AND METHODS： Expression vector pcDNA3-HBx was constructed by recombination DNA technique and tmnsfected into HeLa cells using FuGENE6. Expression of x gene in HeLa cells was assessed by Western blotting. Apoptosis in HeLa-Xs cells and HBx transgenic mice hepatocytes was analyzed by flow cytometry. RESULTS： Expression vector pcDNA3-HBx, which could express hepatitis B virus X protein in HeLa cells was constructed. The apoptotic percent in HeLa-Xs cells （9.8%） was higher than blank HeLa cells （0.4% and HeLa-Vs cells（1.3%）. The HBx transgenic mice hepatocytes also showed higher apoptotic percent（54.4% and 40.4% ） than normal mice hepatocytes（6.8% ）. CONCLUSION： Hepatitis B virus X protein could induce apoptosis m vivo and in vitro.