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

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.