乳腺癌阿霉素耐药细胞株MCF-7/ADM的上皮间质化现象及机制  

The EMT and mechanism of human breast cancer cell line MCF-7/ADM

作　　者：冀峰 

机构地区：[1]开封市中心医院乳腺甲状腺外科,河南开封475000

出　　处：《现代肿瘤医学》2017年第20期3225-3228,共4页Journal of Modern Oncology

基　　金：河南省科技厅基础与前沿项目(编号:112300410033)

摘　　要：目的:建立乳腺癌阿霉素耐药细胞株,命名为MCF-7/ADM,探讨耐药细胞的上皮间质化(EMT)现象及机制,为耐药乳腺癌的治疗奠定基础。方法:中等浓度间歇法建立乳腺癌阿霉素耐药细胞株MCF-7/ADM。采用MTT和Transwell实验分别检测耐药对细胞增殖、迁移与转移能力的影响;荧光定量PCR检测MCF-7/ADM细胞EMT相关分子标志物E-钙黏蛋白及间质细胞分子标记N-钙黏蛋白、波形蛋白、纤维连接蛋白等的表达。ELISA检测TGF-β的表达,Western blot检测TGF-β及Smad2/3在蛋白水平的表达。结果:乳腺癌阿霉素耐药细胞MCF-7/ADM的耐药指数为32.2,对5-氟脲嘧啶、顺铂、环磷酰胺产生交叉耐药性。与MCF-7细胞相比,MCF-7/ADM细胞迁移和转移能力明显增强,E-cadherin表达降低而N-cadherin表达显著增高,细胞上清中TGF-β1表达增加,细胞内TGF-β及磷酸化Smad2/3在蛋白水平表达升高。结论:乳腺癌阿霉素耐药细胞MCF-7/ADM发生EMT现象,可能与经典TGF-β通路激活有关。Objective：To investigate the characteristic and mechanism of the cell migration and invasion ability of breast cancer cells with acquired resistance to chemotherapy. Methods ： To establish a human muhidrug - resistant breast tumor cell line（ MCF -7/ADM） by middle doses of intermittent induction. The biological behavior of MCF -7/ ADM cells examined in the study included cell proliferation, migration and metastasis by MrI and Transwell tests. The mRNA that the MCF -7 and MCF -7/ADM cells expression levels of the selected genes （E -cadherin, N -cadherin, Vimentin,Fibronectin） were validated with real- time quantitative PCR. To detect the TGF-β of the serum by ELISA, to detect the expression of TGF - β and p - Smad2/3 in cells by Western blot. Results ： The drug resistant in- dex of MCF - 7/ADM cell kept on 32.2 times over MCF - 7 cell. MCF - 7/ADM cells had obvious cross - resistance to other chemotherapy drugs. The ability of cell growth rate, migration and invasion was enhanced significantly in MCF - 7/ADM compared to MCF cells. MCF - 7/ADM cells expressed increased levels of mRNA for N - cadherin but de- creased the level of E - eadherin. MCF - 7/ADM cells expressed higher TGF - βand p - Smad2/3 contrast to MCF - 7 cells. Conduslon： MCF - 7/ADM ceils exhibited epithelial - mesenchymal transition phenotype which is related to TGF - β pathway.

关 键 词：乳腺癌 耐药 阿霉素 上皮间质化 机制 

分 类 号：R737.9[医药卫生—肿瘤]