CHO细胞株中高危型HPV-16E6与低危型HPV-11E6的定位分析  

Location and expression of high-risk HPV-16E6 and low-risk HPV-11E6 in CHO cell line

作　　者：孙丽娜[1] 申辛欣[2] 魏建春[1] 张慧娟[1] 张恩民[1] 马凤琴[1] 

机构地区：[1]传染病预防控制国家重点实验室中国疾病预防控制中心传染病预防控制所,北京102206 [2]传染病预防控制国家重点实验室中国疾病预防控制中心病毒病预防控制所,北京100050

出　　处：《中华肿瘤防治杂志》2010年第3期173-176,共4页Chinese Journal of Cancer Prevention and Treatment

摘　　要：目的:探讨高危型HPV-16E6与低危型HPV-11E6生物学行为的差异。方法:构建真核表达载体pGFP-16E6和pGFP-11E6,转染CHO细胞,荧光显微镜下动态观察E6蛋白的定位和表达水平。结果:CHO细胞在转染pGFP质粒后的21h,GFP表达到达高峰,高危型GFP-16E6主要定位于细胞核内,低危型GFP-11E6主要定位于细胞质内,与GFP均匀分布于整个细胞明显不同;自转染后6h,GFP和GFP-16E6蛋白开始表达(荧光强度为54.12和48.27),而GFP-11E6蛋白在转染后的12h才开始表达(85.64),至21h时,GFP、GFP-16E6和GFP-11E6蛋白的表达均达到高峰(132.19、121.94和127.91),P<0.05;以后,随时间的延长蛋白表达逐渐降低。结论:HPV-16E6主要定位于细胞核,HPV-11E6主要定位于细胞质,这或许可以成为它们致病性差异的一种解释。OBJECTIVE：To investigate the difference of biological behavior between high-risk HPV-16E6 and low-risk HPV-11E6 proteins.METHODS：Eeukaryotic expression plasmids named pGFP-16E6 and pGFP-11E6 were constructed,then tranfected to CHO cell line and the E6＇s location was observed by a fluorescent microscope dynamically.RESULTS：GFP reached the highest expression level at 21 h post-transfection.After transfecting,GFP-16E6 mainly expressed in the nuclei of the three cells used,but GFP-11E6 exclusively in the cytoplasm of all the cells used.The results indicated that GFP and GFP-16E6 proteins were expressed from 6 h post-transfection （54.12,48.27）.While GFP-11E6 protein was not detected at 6 h post-transfection,it expressed from 12 h post-transfection （85.64）.The expressions of the three proteins increased gradually,and reached their maximum expression levels at 21 h （132.19,121.94,127.91,〈P0.05）.Then,they decreased gradually.CONCLUSION：HPV-16E6 mainly expresses in the nuclei,but HPV-11E6 exclusively in the cytoplasm,and this may be one of the reasons why only the high-risk HPV E6 is associated with cancer.

关 键 词：乳头状瘤病毒 人 绿色荧光蛋白质类 CHO细胞 

分 类 号：R450[医药卫生—治疗学]