幽门螺杆菌GGT基因在胃癌上皮细胞中的表达和定位  

Expression and subcellular localization of Helicobacter pylori GGT gene in SGC-7901 cells

作　　者：唐炜[1] 尹江涛[1] 张尤历[1] 

机构地区：[1]江苏大学附属医院,江苏省镇江市212001

出　　处：《世界华人消化杂志》2015年第18期2880-2885,共6页World Chinese Journal of Digestology

摘　　要：目的:克隆幽门螺杆菌(Helicobacter pylori,H.pylori)γ-谷氨酰转肽酶(γ-glutamyl transpeptidase,GGT)基因,实现GGT基因在人胃癌上皮细胞SGC-7901中的表达和定位.方法:本实验从胃癌患者胃黏膜组织中分离培养获得H.pylori,提取其基因组DNA,对GGT基因进行PCR扩增,将利用绿色荧光蛋白(green fluorescent protein,GFP)融合GGT基因全长序列和成熟肽序列,克隆进pcDNA3质粒中,观察GGT基因表达后在人胃癌上皮细胞SGC-7901中的定位,观察SGC-7901细胞形态、生长以及运动性的变化.推测其可能诱导细胞凋亡的途径.结果:pcDNA3-GGT-EGFP质粒转染SGC-7901细胞后,在SGC-7901细胞中表达外源基因,在显微镜下可见转染的细胞产生明显的病变,细胞由梭形变成圆形、变大、胞核扩大.转染60 h后,在共聚焦显微镜下观察,发现荧光信号主要集中在细胞质中.结论:成功克隆了GGT基因,经酶切和测序验证正确,成功构建了pcDNA3-GGT-EGFP质粒.AIM：To investigate the expression and subcellular localization of Helicobacter pylori（H.pylori） γ-glutamyl transpeptidase（GGT） gene in SGC-7901 cells.METHODS：H.pylori strains were isolated from the gastric mucosa of patients with gastric cancer,and cultured on solid agar medium.The GGT gene was amplified from the chromosomal DNA of H.pylori by PCR,and was cloned into T vector.The recombinantplasmids were confirmed by enzyme digestion and sequence analysis.H.pylori GGT was then expressed and localized in SGC-7901 cells.RESULTS：The GGT gene was obtained and its sequence was proved to be correct by sequencing identification.SDS-PAGE and Western blot analysis showed that the GGT product could be expressed in SGC-7901 cells.CONCLUSION：The GGT gene has been successfully expressed in SGC-7901 cells and localized in the cytoplasm.This lays a foundation for studying the relationship between GGT and mitochondria-mediated apoptosis.

关 键 词：幽门螺杆菌 GGT基因 细胞表达 定位 

分 类 号：R735.2[医药卫生—肿瘤]