人巨细胞病毒感染对宿主细胞周期蛋白表达影响的实验研究  

Experimental study of the effect of human cytomegalovirus infection on the expression of cyclins

作　　者：刘楠[1] 余艳红[1] 钟梅[1] 闻良珍[2] 

机构地区：[1]南方医科大学南方医院妇产科,广州510515 [2]华中科技大学同济医院附属同济医院妇产科,武汉430030

出　　处：《中国优生与遗传杂志》2007年第5期26-27,共2页Chinese Journal of Birth Health & Heredity

基　　金：国家自然科学基金资助项目(项目批准号30271673)

摘　　要：目的观察人巨细胞病毒(HCMV)感染对宿主细胞周期蛋白(Cyclins)表达的影响。方法用HCMV感染同步化于G0/G1期的人胚肺成纤维细胞(HEL),分别于感染后0 h、3 h、6 h、12 h、24 h、48 h、72 h、96 h终止培养。用免疫蛋白印迹法(Western Blot)检测CyclinE、CyclinA、CyclinD1蛋白的表达。结果HCMV感染接触抑制细胞12 h时CyclinE蛋白被诱导,感染后24 h出现CyclinE峰值;HCMV不能诱导CyclinA蛋白表达;CyclinD1没有被诱导,且在感染后24 h开始下降。结论HCMV感染同步化于G0/G1期的细胞后,诱导CyclinE蛋白明显升高,激活CyclinE/Cdk2激酶,使细胞周期越过G1/S限制点,将细胞周期阻止于晚G1期。Objective ： To observe the effect of human cytomegalovirus infection on expression of cyclines. Metheds ： HCMV infected cell was established in vitro by incubating passage cultured HEL and HCMV AD169 strain with different titres. The cells were synchronized in the GO/G1 stage by contact inhibition and infected with strain AD169 of HCMV at an MOI of 5 PFU per cell. We harvested infected cells at different time 0h, 3h, 6h, 2h, 24h, 48h, 72h and 96h post infection. Then expression of proteins ofcyclineE, cyclineA and cyclineD1 were determined with Western Blot. Results： CyelinE protein was induced 12h PI and peak induction occurred 24h PI in contact - inhibited cells. CyclinA protein expression was not induced in HCMV infected density - arrested cells. CyclinD1 was not induced by the virus. Rather, the abundanc of cyclinD1 decreased 24 h PI. Conclusion： The expression of cyclinE and activity of cyclinE/Cdk2 kinase are increased obviously in GO/G1 stage cells infected with HCMV, which may induce the cell cycle to overpass G1/S restriction point and make the cell cycle arrested in later G1 stage.

关 键 词：HCMV感染细胞周期蛋白免疫蛋白印迹法 

分 类 号：R373.4[医药卫生—病原生物学]