STGC3基因高表达Daudi细胞系的建立与鉴定  

Establishment and Identifition of Daudi Cell Line Over-expressed STGC3 Gene

作　　者：肖建忠[1] 贺修胜[1] 邱清朝[1] 邓敏[1] 胡波[1] 罗桥[1] 

机构地区：[1]南华大学肿瘤研究所,湖南衡阳421001

出　　处：《中南医学科学杂志》2011年第4期367-371,共5页Medical Science Journal of Central South China

基　　金：国家自然科学基金资助项目(30470967)

摘　　要：目的探讨STGC3基因高表达对Daudi细胞形态、倍增时间和增殖速度的影响。方法采用电穿孔基因转染技术,将pCDNA3.1(+)/STGC3真核表达载体导入Daudi细胞系,G418筛选,建立高表达STGC3基因的Daudi细胞系,利用Western-blotting检测转基因细胞系中STGC3蛋白的表达;运用免疫细胞化学和HE染色方法及绘制细胞生长曲线,了解STGC3基因表达在Daudi细胞中的定位,以及对细胞倍增时间和增殖速度的影响。结果在转基因的Daudi细胞系中,STGC3蛋白高表达并定位于胞浆与胞核。转染STGC3基因后,Daudi细胞倍增时间延长,增殖速度明显减慢(P<0.05)。结论成功建立STGC3基因高表达Daudi细胞系。Objective To investigate celluar morphous,doubing time and proliferation speed of Daudi cells affected by STGC3 gene over-expression.Methods We adopted the method of electroporation gene transfection method to introduc pCDNA3.1（＋） /STGC3 eukaryotic expression plasmid into Daudi cell,which was selected by geneticin,and established the Daudi cell line of STGC3 gene over-expression.Then,we used Western-blotting to detect expression of STGC3 protein in transgenic Daudi cell,and deployed immuocytochemistry,HE staining and cell growth curve methods to observe the locating of STGC3 protein,the celluar doubing time and proliferation speed affected by STGC3 gene over-expression in Daudi cells.Results STGC3 protein over-expressed and located in endochylema and nucelus of transgenic Daudi cell line.The celluar doubing time was extended,and the celluar proliferation speed was stepped down by STGC3 gene over-expression in Daudi cells（n=3,P〈0.05） .Conclusion The Daudi cell line of STGC3 gene over-expression was established successfully.

关 键 词：STGC3基因 高表达 DAUDI细胞 细胞增殖 

分 类 号：R730.26[医药卫生—肿瘤]