携带BARF1基因的人胃上皮细胞株的建立

Establishment of human gastric epithelial cells carried BARF1 gene

目的建立携带BARF1基因的人胃上皮细胞株,为探讨EB病毒编码BARF1基因对胃上皮细胞的影响而建立体外模型。方法构建携带BARF1基因的真核载体pcDNA3.1(+)-his/BARF1,转染人胃上皮细胞系GES-1,G418筛选单克隆。用CCK-8检测未转染的GES-1、转染空载体GES-1及转染BARF1基因的GES-1增殖状况。结果双酶切鉴定,666bp的BARF1基因片段已连接到真核表达载体pcDNA3.1(+)-his。转染细胞通过G418筛选,获得了稳定表达BARF1基因的人胃上皮细胞株,转染BARF1基因的GES-1细胞增殖速度显著高于未转染的GES-1和转染空载体GES-1细胞(P<0.01)。结论建立了稳定表达BARF1基因的人胃上皮细胞株。

The purpose of this study was to establish human gastric epithelial cells carried BARF1 gene,and construction in vitro model for studying the role of BARF1 gene in proliferation of gastric epithelial cells.The eukaryotic vector pcDNA3.1（＋）-his/BARF1 for carried BARF1 gene was constructed and transfected into human gastric epithelial cell line GES-1,monoclonal was selected by G418.The proliferation status of gastric epithelial cells in untransfected gene BARF1,transfected empty vector and transfected BARF1 gene were detected with CCK-8.666bp products of BARF1 had been correctly connected to the eukaryotic expression vector pcDNA3.1（＋）-his through double enzyme digestion.Obtained human gastric epithelial cells of stable expression BARF1 gene by G418 was selected.The proliferation of GES-1 cells transfected BARF1 gene was significantly higher than that of untransfected gene BARF1 and transfected empty vector（P〈0.01）.Results indicated that human gastric epithelial cells carried BARF1 gene were established successfully.