HPV16 E6蛋白与hDaxx的相互作用及其对HeLa细胞凋亡的影响  

Effect of interaction of human papillomavirus type 16 E6 protein and hDaxx on apoptosis of HeLa cells

作　　者：王鑫[1] 朱翠明[1] 刘安元[1] 尹卫国[1] 李金丽[1] 蔡恒玲[1] 万艳平[1] 

机构地区：[1]南华大学病原生物学研究所,衡阳421001

出　　处：《肿瘤》2008年第4期293-296,共4页Tumor

基　　金：湖南省教育厅省属高校科研计划资助项目(编号:02C391)

摘　　要：目的:研究HPV16 E6(human papillomavirus type16 E6)蛋白与hDaxx(human death domain associated protein)的相互作用及其对HeLa细胞凋亡的影响,为探索HPV16 E6蛋白的致癌机制提供实验依据。方法:构建pGADT7-E6和pGBKT7-hDaxx重组载体,利用酵母双杂交系统检测HPV16 E6蛋白与hDaxx的相互作用,并用Western印迹法检测二者在酵母菌中的表达。将E6与hDaxx的真核表达载体共转染HeLa细胞,用5-氟尿嘧啶(5-FU)诱导凋亡,FCM法检测凋亡率。结果:E6蛋白与hDaxx在细胞内存在相互作用。共转染pcDNA3.1(-)/E6和pcDNA3.1(-)/hDaxx的HeLa细胞,随pcDNA3.1(-)/hDaxx转染量的增加凋亡率上升,差异有统计学意义(P<0.01)。结论:HPV16 E6蛋白与hDaxx在细胞内相互作用,hDaxx过表达可提高表达E6蛋白的HeLa细胞对5-FU的敏感性,且这种关系存在剂量依赖性。E6蛋白可能通过和hDaxx的相互作用抑制细胞凋亡。Objective：To study the interaction of human papillomavirus type 16 （HPV16 E6） protein and human death domain associated protein （hDaxx） and its effect on apoptosis of HeLa cells to provide the experimental basis for exploring the oncogenic mechanism of HPV16 protein. Methods： Recombinant vectors of pGADT7/E6 or pGBKT7/hDaxx were constructed. The interaction of E6 protein and hDaxx was detected by yeast two-hybrid system. Their expression in yeast was detected by Western blotting. The eukariotic plasmids of E6 and hDaxx were transfected into HeLa cells. Apoptosis was induced by 5-FU. The apoptotic ratio was measured by flow cytometry. Results：E6 protein had intracellular interaction with hDaxx. The apoptotic rate was rising with the increase in the transfection quantity of pcDNA3.1 （ - ）/hDaxx in pcDNA3.1 （ - ）/E6 and pcDNA3.1 （ - ）/hDaxx co-transfected cells. The difference was significant （P 〈0.01 ）. Conclusion ：There existed an intracellular interaction between HPV16 E6 protein and hDaxx. The over-expression of hDaxx could increase the sensitivity of E6 protein-positive HeLa cells to 5-FU. The effect was in a dose-dependent manner. HPV16 E6 protein inhibited the apoptosis of HeLa cells by interacting with hDaxx.

关 键 词：乳头瘤病毒E6蛋白质类 HELA细胞 双杂交系统技术 细胞凋亡 蛋白hDaxx 

分 类 号：R737.33[医药卫生—肿瘤]