HER-2/neu对雌激素依赖性子宫内膜癌细胞信号通路MAPK/ERK调控的研究  

Regulating effect of HER-2/neu gene in endometrial cancer cells of estrogen-dependent MAPK/ERK signal pathway

作　　者：于志娟[1] 马晓欣[1] 张茹[1] 柯孝瑜[1] 

机构地区：[1]中国医科大学附属盛京医院妇产科,辽宁沈阳110004

出　　处：《现代肿瘤医学》2013年第8期1684-1688,共5页Journal of Modern Oncology

基　　金：国家自然科学基金资助项目(编号:81272874);辽宁省教育厅科技基金项目(编号:L2010642);辽宁省科学技术厅科技基金项目(编号:2011404013-8);沈阳市科技局科技基金项目(编号:F12-227-1-62)

摘　　要：目的:探讨HER-2/neu对雌激素依赖性子宫内膜癌细胞信号通路MAPK/ERK的调控机制。方法:EGF(表皮生长因子)处理Ishikawa细胞株及转染HER-2/neu的Ishikawa细胞株,Western blot检测转染前后细胞中COX-2、p-ERK/ERK蛋白的表达。ELISA方法检测细胞培养上清液中雌二醇的含量。用MAPK/ERK的抑制剂PD98059抑制信号通路,分别以50μmol/L,于不同时间(5、10、20、30、60min)及不同浓度(5、10、50、100μmol/L)作用30min后,Western blot检测COX-2的表达水平、ELISA检测雌二醇水平。结果:转染HER-2/neu的Ishikawa细胞株COX-2、p-ERK/ERK的蛋白表达量及细胞上清液中雌二醇的含量明显高于未转染的Ishikawa细胞株(P<0.05)。应用PD98059抑制MAPK/ERK通路,随PD98059浓度的增加及作用时间的延长,两组细胞COX-2及雌二醇的表达均逐渐减少,且抑制作用与浓度及作用时间呈依赖关系;但转染组COX-2与E2降幅大于未转染组(P<0.05)。结论:HER-2/neu可能通过MAPK/ERK两条通路来诱导COX-2的转录,进而导致雌激素的分泌增多,使细胞无限生长。Objective：To explore the effect of HER-2/neu gene in endometrial cancer cells of estrogen-dependent MAPK/ERK signal pathway.Methods：EGF stimulated Ishikawa cells,Ishikawa cells stably transfected with HER-2/neu plasmid,Western blotting was used to detect transfected cells before and after COX-2,p-ERK/ERK protein expression levels,ELISA to detect transfected cell culture supernatant before and after the content of E2.The blockage of MAPK/ERK pathway PD98059 inhibits pathway,50μmol/L for 5min,10min,20min,30min and 60 min;5μmol/L,10μmol/L,50μmol/L and 100μmol/L for 30 min,Western blotting was used to detect transfected cells before and after COX-2 protein expression levels,ELISA to detect transfected cell culture supernatant before and after the content of E2.Results：COX-2,p-ERK/ERK,E2 protein expression in HER-2/neu plasmid transfection group was significantly higher than control group（P 0.05）;The results showed a long with increasing drug conce-ntration and prolonging action time,COX-2,E2 protein expression in each group was gradually decreased,the inhibitive effect has positive correlation with the concentration and action time.Conclusion：HER-2/neu promotes the growth of Ishikawa cells by regulating the MAPK/ERK pathway to affect the expression of COX-2,E2 in vivo.

关 键 词：子宫内膜癌 HER-2 neu(c-erbB-2) MAPK ERK通路 雌激素 

分 类 号：R730.23[医药卫生—肿瘤] R737.33[医药卫生—临床医学]