小鼠α1,3-半乳糖基转移酶基因转染胃癌GC9811细胞的功能研究  

GASTRIC CANCER CELL GC9811 TRANSFECTED BY MURINE α1,3-GALACTOSYLTRANSFERASE GENE

作　　者：郭军[1] 丁杰[1] 喻召才[1] 李韧[1] 樊代明[1] 

机构地区：[1]西安第四军医大学西京医院

出　　处：《解放军医学杂志》2003年第4期316-318,F003,共4页Medical Journal of Chinese People's Liberation Army

摘　　要：通过RT PCR从BALB/c小鼠腹腔巨噬细胞扩增出编码小鼠α1,3 半乳糖基转移酶基因的全长cDNA并构建其真核表达载体pCDNA3 1/V5 HisAα1,3GT ,pCDNA3 1/V5 HisAα1,3GT通过脂质体转染胃癌GC9811细胞 ,使胃癌细胞表达Galα(1,3)Gal糖表位 ,G4 18筛选 2个月内的阳性克隆 ,RT PCR鉴定转染细胞 ,间接免疫荧光染色检测转染真核表达载体的癌细胞Galα(1,3)Gal的表达 ,利用人体血清内针对Galα(1,3)Gal糖表位的天然抗体对癌细胞进行杀伤。结果表明 ,成功构建了α1,3 半乳糖基转移酶基因真核表达载体 ,转染α1,3 半乳糖基转移酶基因的癌细胞高表达Galα(1,3)Gal糖表位 。The full length cDNA of α1,3 galactosyltransferase gene was amplified by RT PCR from the peritoneal macrophages of BALB/c mice, which was subcloned into an eukaryotic expression vector pCDNA3 1/V5 HisA, and it was confirmed by DNA sequencing. The gastric cancer cell GC9811 was transfected with pCDNA3 1/V5 HisAα1,3GT by Lipofectamine TM 2000. The positive clone was screened by G418 for two months and confirmed by RT PCR. The gastric cells expressing Galα(1,3)Gal epitope were detected by immuohistochemisty, and they were killed by naive antidoby specific to Galα(1,3)Gal epitope. The results showed that the pCDNA3.1/V5 HisAα1,3GT was successfully constructed, and the transfected gastric cancer cells could highly express Galα(1,3)Gal, which promoted lysis of the cancer cells by normal human serum.

关 键 词：小鼠 α1 3—半乳糖基转移酶基因 转染 胃癌 RT—PC 间接免疫荧光染色 

分 类 号：R735.2[医药卫生—肿瘤]