高危型HPV-16E6在小鼠骨髓源性树突细胞的表达  

Srudy on location and expression of high-risk HPV-16E6 in dentritic cell

作　　者：王淑京[1] 黄元铭[1] 温晓婷[1] 孙丽娜[1] 

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

出　　处：《中国热带医学》2013年第4期398-399,416,共3页China Tropical Medicine

基　　金：国家自然科学基金资助(No.81201273)

摘　　要：目的探讨高危型HPV-16E6在树突细胞中的定位。方法构建真核表达载体pGFP-16E6,转染小鼠骨髓源性的树突细胞,在荧光显微镜下动态观察高危型HPV-16E6蛋白在树突细胞内的定位和表达水平。结果树突细胞在分别转染pGFP-16E6和pEGFP-C1质粒后,GFP-16E6主要定位于细胞核内,而对照组的只表达GFP的蛋白则均匀分布于树突细胞。蛋白表达水平的检测表明,在转染树突细胞后的12h,GFP-16E6和GFP蛋白开始表达(荧光强度分别为31.29,39.52),转染后至24h,蛋白表达均到达高峰(荧光强度分别为119.37,134.23),24h后,蛋白表达逐渐降低。结论 HPV-16E6主要定位于树突细胞核内,随时间其蛋白表达水平不同,这为HPV E6的树突细胞疫苗发挥有效的抗癌作用提供理论依据。Objective To investigate the location of high-risk HPV-16E6 protein in dentric cells and the expression of HPV-11E6 fusion protein in mammalian GFP expression system in bone marrowed mouse dentritic cells,to trace the traffic of E6s.Methods Eukaryotic expression plasmids named pGFP-16E6 was coonstructed,then tranfected it into dentritic cells and observed the E6s location by fluorescent microscope dynamically.Results GFP reached the highest expression level 24 hours after transfection.After transfecting,GFP-16E6 mainly expressed in the nuclei of DCs,but GFP control DC expressed in the cytoplasm and nuclei of all the cells.GFP-16E6 and GFP protein were expressed 12 hours after transfection（31.29,39.52）.The proteins expression increased gradually,and reached their maximum expression level at 24 hours（119.37,134.23）.Then it decreased gradually.Conclusions GFP-16E6 mainly expressed in the nuclei of dentric cells,this might explain why the high-risk HPV E6 is associated with cancer.

关 键 词：HPVE6 GFP 定位 蛋白表达 

分 类 号：R512.91[医药卫生—内科学]