HA-CD44st-TGF-β信号通路对乳腺癌MCF-7细胞HER2表达及侵袭能力的影响  

Influence of HA-CD44st-TGF-β signaling pathway on the expression of HER2 and the invasion capability of MCF-7 cell

作　　者：耿鹤 陆颖芝 陈谦[2] 徐静 严海翠 马兆明 黄关宏[2] 房新建[2] GENG He;LU Yingzhi;CHEN Qian;XU Jing;YAN Haicui;MA Zhaoming;HUANG Guanhong;FANG Xinjian(Postgraduate School,Bengbu Medical College, Bengbu 233030,Anhui,China;Department of Oncology,Lianyungang Hospital Affiliated to Bengbu Medical College,Lianyungang 222023,Jiangsu,China)

机构地区：[1]蚌埠医学院研究生院,安徽蚌埠233030 [2]蚌埠医学院附属连云港医院/连云港市第二人民医院肿瘤科,江苏连云港2220230

出　　处：《癌症进展》2019年第7期769-775,共7页Oncology Progress

基　　金：江苏省"333工程"培养资金资助项目(BRA2016299);江苏省卫生和计划生育委员会面上项目(H2017040);连云港市科技局课题资助项目(SH1415)

摘　　要：目的探讨HA-CD44st-TGF-β信号通路对人乳腺癌细胞株MCF-7细胞HER2表达及侵袭能力的影响。方法应用脂质体转染法将真核表达载体pcDNA3.1-CD44st转染入MCF-7细胞中,以该转染细胞株(MCF-7/CD44st)为基础,采用逆转录聚合酶链反应(RT-PCR)、流式细胞术及Western blot检测mRNA及相关蛋白的表达,电泳迁移率变动分析(EMSA)检测激活蛋白-1(AP-1)的活性,Transwell法检测HA-CD44st→TGF-β信号通路对MCF-7细胞侵袭能力的影响。结果 MCF-7细胞转染pcDNA3.1-CD44st后,CD44 mRNA和CD44蛋白的表达水平升高;透明质酸(HA)处理后,MCF-7/CD44st细胞中人表皮生长因子受体2(HER2)、转化生长因子-β(TGF-β)、磷酸化蛋白激酶B(p-AKT)表达水平明显升高,转录因子AP-1活性增强,侵袭细胞数量增多。结论 HA与CD44st受体结合后,可以通过HA-CD44st→TGF-β信号转导通路调节HER2基因表达并增强MCF-7细胞的侵袭能力。Objective To investigate the influence of HA-CD44st-TGF-β signaling pathway on the expression of HER2 and the invasion capability of human breast cancer cell line MCF-7.Method The eukaryotic expression vector pcDNA3.1-CD44st was transfected into MCF-7 cells by Lipofectamine,establishing the transfected cell line MCF-7/ CD44st,which was employed to detect the expression of mRNA and related proteins using reverse transcription- polymerase chain reaction (RT-PCR),flowcytometry,and Western blot,besides,the activity of activator protein-1 (AP-1) and invasion capability of MCF-7 cells were evaluated with electrophoretic mobility shift assay (EMSA) and Transwell assay, respectively.Result The CD44 mRNA and protein were highly expressed in MCF-7 cells transfected with pcDNA3.1- CD44st.The expression of HER2,TGF-β,and p-AKT were markedly elevated,and the activity of AP-1 was promoted, additionally the invasion capability of the cancer cells were increased significantly in MCF-7/CD44st cells treated with hyaluronic acid (HA).Conclusion Binding of HA to CD44st receptor may regulate the expression of HER2 gene and increase the invasion capability of MCF-7 cells through HA-CD44st-TGF-β signaling pathway.

关 键 词：乳腺癌 分化抗原簇44st 转化生长因子-Β HER2 侵袭 

分 类 号：R737.9[医药卫生—肿瘤]