HBXIP基因对乙肝病毒X蛋白诱导细胞凋亡的影响  

Effect of Hepatitis B X Interacting Protein on Apoptosis Induced by Hepatitis B Virus X Protein

作　　者：张晓东[1] 马宏涛[1] 叶丽虹[1] 东楠[1] 史志蔚[1] 沙丽[1] 王洪辉[1] 

机构地区：[1]南开大学生命科学学院,天津300071

出　　处：《中国生物化学与分子生物学报》2005年第3期403-407,共5页Chinese Journal of Biochemistry and Molecular Biology

基　　金：天津市科技发展计划科技攻关重点项目(No.043114311)~~

摘　　要：探讨乙型肝炎病毒X蛋白结合蛋白(hepatitisBXinteractingprotein ,HBXIP)基因在乙型肝炎病毒X蛋白(HBX)诱导肝癌细胞凋亡时对细胞周期的影响.构建HBXIP基因真核表达载体pcDNA3 hbxip ,进行瞬时基因转染,将克隆有HBx基因的pCMV X (分别为1μg、2 μg和3μg)和pcDNA3 hbxip质粒分别和共转染至人H74 0 2肝癌细胞中(总体积分别为5 0 μl) .发现瞬时转染3μgpCMV X质粒后,肝癌细胞凋亡发生率为34 4 % ,肝癌细胞的细胞周期相关蛋白p2 7表达水平发生明显上调;与对照组相比,瞬时转染1μg、2 μg和3μg时,细胞周期蛋白D和细胞周期蛋白E的表达水平均发生明显上调,但随着HBX水平的增加细胞周期蛋白D和细胞周期蛋白E的表达水平发生明显下降;在稳定转染pCMV X质粒的H74 0 2 X肝癌细胞中无明显的细胞凋亡发生,研究发现p2 7的表达水平发生了明显下调,而细胞周期蛋白D和细胞周期蛋白E的表达水平发生了明显上调;当pcDNA3 hbxip质粒与pCMV X质粒进行共瞬时转染时,细胞凋亡发生率由pcDNA3质粒与pCMV X质粒共转染时的2 9 2 %下降为13 3% ,p2 7的表达水平发生了下调,但细胞周期蛋白D和细胞周期蛋白E的表达水平无明显变化.研究结果表明,瞬时转染一定剂量的x基因可导致肝癌细胞发生凋亡,细胞周期相关蛋白p2 7。The aim of this study is to investigate the effect of hepatitis B interacting protein (HBXIP) on apoptosis induced by hepatitis B virus X protein (HBX) in hepatoma cells. The plasmids of pCMV-X (1 μg, 2 μg and 3 μg) and pcDNA3-hbxip were transiently transfected into H7402 cells, a human hepatoma cell line, after successful cloning of pcDNA3-hbxip vector inserted HBXIP gene. The results showed that the apoptosis rate was 34.4% examined by flow cytometery and expression level of p27 was up-regulated by Western blot analysis when 3μg pCMV-X plasmid was transfected. Comparison with control group, expression level of cyclin D and cyclin E were up-regulated when 1 μg, 2 μg and 3 μg pCMV-X plasmids were trasnfected, respectively, but gradually decreased following the increasing expression level of HBx. However, the expression of p27 was down-regulated and expression of cyclin D and cyclin E was up-regulated in H7402-X cells without apoptosis, that were HBV x gene stable transfection cells. The apoptosis rate was 13.3%, and the expression level of p27 was down-regulated when both pCMV-X and pcDNA3-hbxip plasmids were transiently co-transfected into H7402 cells. But no change of expression level of cyclin D and cyclin E were found. In conclusion, our data suggest that HBx is able to induce apoptosis in hepatoma cells.The proteins regulating cell cycle,such as p27,cyclin D and cyclin E, were involved in the apoptosis.HBXIP protein can inhibit the apoptosis induced by HBX.

关 键 词：肝癌 乙型肝炎病毒X蛋白 乙型肝炎病毒X蛋白结合蛋白 细胞周期 细胞凋亡 

分 类 号：R364.7[医药卫生—病理学] Q255[医药卫生—基础医学]