幽门螺杆菌对人胃癌细胞SGC-7901中SHP-2及细胞骨架的影响  

Influences of Helicobacter pylori on SHP-2 expression and cytoskeleton in human gastric cancer cells SGC-7901

作　　者：张尤历[1] 陆芬英[1] 王文兵[2] 张宇川[1] 刘勇攀[1] 吴莺[1] 

机构地区：[1]江苏大学附属医院消化内科,江苏省镇江市212001 [2]江苏大学生命科学研究院,江苏省镇江市212013

出　　处：《世界华人消化杂志》2008年第26期2916-2921,共6页World Chinese Journal of Digestology

基　　金：江苏省社会发展资助项目;No.BS2006041~~

摘　　要：目的:研究幽门螺杆菌(Helicobacter pylori,H pylori)刺激人胃癌细胞SGC-7901后,对细胞内SHP-2表达及细胞骨架的影响.方法:将临床分离的胃癌H pylori菌体裂解,以超声提取物刺激SGC-7901细胞1h,逆转录聚合酶链式反应(RT-PCR)半定量法测定刺激前后SHP-2表达量的变化;并将PCR产物进行克隆和测序.将不同疾病(胃炎,胃溃疡,胃癌)的菌株H pylori超声提取物刺激SGC-7901细胞10min,采用phalloidin对细胞内骨架进行荧光染色,研究其对细胞骨架的影响.将野生型和突变型RhoA质粒转入细胞后,再以胃癌H pylori超声提取物刺激,进一步观察细胞内骨架的变化.结果:胃癌H pylori菌体超声提取物刺激SGC-7901细胞1h后,细胞内SHP-2表达量没有明显变化.胃炎,胃溃疡,胃癌H pylori菌体超声提取物刺激人胃癌细胞SGC-7901后,与对照组相比,细胞内骨架增多(65.4±0.510,63.37±0.475,64.53±0.522vs20.34±1.376,均P<0.05),但这三种菌株间相比无统计学意义.转染入野生型RhoA质粒的细胞经胃癌H pylori菌体超声提取物刺激后,细胞内骨架较未转染者增多(72.99±1.818vs61.78±1.288,P<0.05),而转染入无活性突变型RhoA质粒的细胞经刺激后,细胞内骨架与未转染者无明显差别.结论:胃癌H pylori超声提取物未能引起细胞内SHP-2mRNA表达量的改变;胃炎,胃溃疡,胃癌H pylori菌体超声提取物能够增加细胞内骨架的形成,其作用与H pylori菌种无关,RhoA活性的增强是其中的一个重要中间环节.AIM：To investigate the changes of SHP-2 expression and cytoskeleton in human gastric cancer SGC-7901 cells stimulated by Helicobacter pylori（H pylori）. METHODS：H pylori was isolated from human gastric mucosa and cultured on solid culture plate.The SGC-7901 cells were incubated with the sonicating extract from gastric cancer for 1 h, and the mRNA expression of SHP-2 was measured using semi-quantitative PCR.The PCR products were then cloned and sequenced.After stimulating the SGC-7901 cells with the sonicat- ing extract from gastritis,gastric ulceration and gastric cancer for 10 min,we observed the cy- toskeleton using F-actin phalloidin fluorescent staining.After the wild type and mutant pcDNA RhoA was transfected into the cells and stimulated with the sonicating extract from gastric cancer H pylori,the cytoskeleton was further observed. RESULTS：There was no remarkable change in mRNA expression of SHP-2 following stimulation of SGC-7901 cells with gastric cancer H pylori sonicating extract.After being compared with the sequences in GenBank,the PCR products proved to be SHP-2.However,after stimulation with either gastritis,gastric ulcer and gastric cancer H pylori sonicating extract,the cytoskeleton in SGC-7901 cells significantly increased compared with the control group（65.4±0.510, 63.37±0.475,64.53±0.522 vs 20.34±1.376,all P 〈0.05）,but there were no significant difference among the three groups.After stimulation with gastric cancer H pylori sonicating extract,the cytoskeleton in cells transfected with wild type pcDNA RhoA were increased compared with the untransfected cells（72.99±1.818 vs 61.78± 1.288,P〈0.05）;while mutant pcDNA RhoA had no effect on the cytoskeleton. CONCLUSION：The sonicating extract from gastric cancer H pylori has no effect on mRNA expression of SHP-2 in human gastric cancer SGC-7901 cells;however sonicating extract from gastritis,gastric ulcer and gastric cancer can in- crease the cytoskeleton in SGC-7901 cells,and this function is independent of the category of

关 键 词：幽门螺杆菌 胃癌细胞 SHP-2 细胞骨架 RHOA 

分 类 号：R735.2[医药卫生—肿瘤]