EB病毒感染对人胃上皮细胞GES-1增殖的影响

目的:探讨EB病毒感染人胃上皮细胞GES-1后对GES-1增殖的影响。方法:用携带新霉素抗性(NEOr)基因的重组EBV以密切接触法感染人胃上皮细胞系GES-1,同时以正常人胃上皮细胞GES-1为对照。经G418筛选出EBV感染阳性克隆,进行细胞形态学检测,并采用免疫组织化学染色法测定EBNA1的表达以鉴定EBV感染细胞克隆。对上述细胞用细胞计数法及MTT法检测细胞增殖情况;流式细胞术对细胞周期进行分析。结果:感染EBV的GES-1细胞大部分为EBNA1表达阳性,通过G418筛选,获得了稳定的EBV感染细胞克隆。EBV感染组细胞体积变大,核浆比增加。细胞计数法及MTT法绘制细胞生长曲线显示细胞增殖速度明显加快。流式细胞分析结果显示EBV感染组细胞S期延长。结论:EBV感染后使GES-1细胞稳定且高表达EBNA1,并且能够促进GES-1细胞的增殖。

Effect of EBV Infection on the proliferation of Gastric Epithelial Cell GES-1

Objective:To analyze the relationship between the infection of EBV and the proliferation of human gastric epithelial cell GES-1.Methods:In this experiment,recombined EBV Akata 1061 carrying neomycin resistance gene(NEOr)was used to infect GES-1 by intimate contact method,in the mean time the normal GES-1 cell was used as control.And then,the geneticin 418(G418)was adopted to select positive clones in EBV infected cells.The morphologic changes of GES-1 cells were observed under optical microscope.The method of immunohistochemical(IHC)staining was used to detect EBNA1 expression in EBV infected cell clones.Cell counting and MTT method were adopted to detect the growth of cell.And flow cytometry(FCM)was adopted to measure the changes of cell cycle progress.Results:Stable cell clones were obtained by EBV infectd after G418 selecting.Compared with the control group,immunohistochemical staining showed that EBV infected cells express EBNA1 much more obviously.Compared with the control groups,the volume of cells and karyo-plasmic ratio were increased after infectd by EBV.The results of cell counting and MTT showed that the growth of EBV infected cells was faster than that of un-infected cells.The result of FCM analysis showed that the proportion of S-phase cells in EBV infected group increased obviously.Conclusion:The infection of EBV makes the cells of GES-1 express EBNA1 steadily and highly,and enhances the proliferation of GES-1.