高危型HPV-16E6在小鼠骨髓源性树突细胞的表达 |
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引用本文: | 王淑京,黄元铭,温晓婷,孙丽娜. 高危型HPV-16E6在小鼠骨髓源性树突细胞的表达[J]. 中国热带医学, 2013, 13(4): 398-399,416 |
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作者姓名: | 王淑京 黄元铭 温晓婷 孙丽娜 |
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作者单位: | 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 |
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基金项目: | 国家自然科学基金资助(项目编号:81201273) |
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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。
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关 键 词: | HPVE6 GFP 定位,蛋白表达 |
Study on location and expression of high-risk HPV-16E6 in dentritic cell |
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Affiliation: | WANG Shu-jing,HUANG Yuan-pei,WEN Xiao-ting,et al.(State key Laboratory for Infectious Disease Prevention and Control,National Institute for Infectious Disease Control and Prevention;Chinese Center for Disease Control and Prevention,Beijing 102208,P.R.China) |
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Abstract: | 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. |
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Keywords: | HPV E6 GFP Location Protein expression |
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