EB病毒转化人胃上皮细胞GES-1中bcl-2基因的表达

目的 研究EBV感染人胃上皮细胞系GES-1后bcl-2基因的表达状况,探讨6cl-2基因在EBV相关胃癌发生中所起的作用.方法 采用携带新霉素抗性(NEOr)基因的重组EBV产生细胞系Akata 1061以密切接触法感染人胃上皮细胞系GES-1,经有限稀释法克隆和G418筛选获得感染和转染细胞克隆,用pcDNA3空载体质粒转染的人胃上皮细胞系GES-1细胞作对照:采用免疫细胞化学染色(ICC)法测定EBNA1基因的表达以鉴定EBV感染细胞克隆及测定EBV感染细胞中Bcl-2蛋白的表达.结果 与对照相比较,感染细胞中Bcl-2蛋白的表达阳性.结论 EBV感染可使人胃上皮细胞Bcl-2蛋白表达增高,EBv对人胃上皮细胞的转化作用可能与bcl-2基因的过度表达有关.

Expression of bcl-2 gene in EBV-transformed human gastric epithelial cell line GES-1

Objective To detect bcl-2 gene expression in Epstein-Barr virus (EBV)-infected human gastric epithelial cell line GES-1 for understanding the role of bcl-2 gene in the carcinogenesis of EBV-associated gastric carcinoma. Methods Akata 1061 cells producing recombined EBV carrying neomycin resistance gene (NEOr) was used to mediate the EBV infection of human gastric epithelial cell line GES-1 via close contact, with the empty plasmid pcDNA3-transfected GES-1 cells via lipofectamine method as a control. The EBV-infected and pcDNA3-transfected cells were cloned by limited dilution and the positive clones selected with G418. Immunocytochemical staining was performed to detect the expressions of EBNA1 and Bcl-2 protein. Results Bcl-2 protein expression was detected in EBV-infected cells but not in the control cells. Conclusion EBV infection can increase Bcl-2 expression in gastric epithelial cells, and such cell transformation effect of EBV is related to the overexpression of bcl-2 gene.