人巨细胞病毒感染对体外培养成纤维细胞E2F1表达的时序性影响  

The sequential effect of human cytomegalovirus infection on the expression of E2F1 in cultured human embryo lung fibroblast in vitro

作　　者：刘志峰[1] 方峰[1] 甄宏[1] 李革[1] 董永绥[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院儿科学系

出　　处：《中华微生物学和免疫学杂志》2005年第9期707-710,共4页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金资助项目(批准号:30070928)

摘　　要：目的研究人巨细胞病毒(human cytomegalovirus,HCMV)感染对体外培养成纤维细胞E2F1表达的时序性影响,进一步揭示HCMV的致病机制.方法用高和低感染复数(multiplicity of infection,MOI)HCMV病毒感染体外培养的成纤维细胞,在不同时间点用Western blot法检测细胞E2F1蛋白表达水平.结果低MOI和高MOI感染都可时序性上调宿主细胞E2F1蛋白表达,均以感染后2h上调幅度最高(2～3倍).高MOI感染的上调作用强于低MOI感染.结论HCMV在感染初期即可激发宿主细胞E2F1表达明显增加,并在24h内呈时序性上调,提示E2F1在病毒感染早期诱导宿主细胞向S期和G2/M期偏移,进而在营造有利于病毒复制微环境的机制中起关键作用.Objective To investigate the time-sequential effect of HCMV infection on the expression of E2FI in human embryo lung fibroblast （HELF） culture in vitro in order to elucidate the mechanism of HCMV infection. Methods The human embryo lung fibroblasts were challenged with high MOI and low MOI of HCMV AD169 strain, respectively, the normal cells served as controls. The expressing level of E2F1 in each group of the cultures was measured by Western blot at 2 h, 4 h, 6 h, 8 h, 12 h, 16 h, 20 h, 24 h, 48 h, 72 h and 96 h after infection. Results Both high and low MOI of HCMV infection could up-regulate the expression of E2F1 in time- sequence within the first 24 h after infection. The expression peak appeared at 2 h after infection （2-3 times）, and the up-expressing effect of high MOI infection was higher than that of low MOI infection. Conclusion E2F1 ex- pression of host cells could be markedly induced in the early period after HCMV infection, and display a time-sequential up-regulation within the first 24 h. It was proved thai E2F1 transcriptional factor plays an important role in the drift of cells to S and G2/M phases during the early period of infection which may be beneficial to the reproduction of HCMV virttses.

关 键 词：人类巨细胞病毒 E2F 转录活性 人巨细胞病毒感染 成纤维细胞 E2F1 体外培养 时序性 蛋白表达水平 Western 宿主细胞 上调作用 

分 类 号：R373[医药卫生—病原生物学]