乳腺癌间质细胞对MCF-7细胞Wnt1、Notch 1、β-catenin表达及细胞迁移的影响

Effects of breast cancer stromal cells on expression of Wnt1,Notch1 and β-catenin and migration of MCF-7 cells

目的研究乳腺癌间质细胞对乳腺癌细胞的调控效应,探讨微环境在乳腺癌发生和发展中的作用。方法取5例乳腺癌患者手术后标本进行原代间质细胞的分离、培养和纯化,并行免疫组化鉴定。采用Transwell三维共培养体系,将原代间质细胞与乳腺癌细胞系(MCF-7)共培养3d(实验组),以2%FCS培养基单独培养MCF-7作为对照组。Real-time PCR检测MCF-7的Wnt1、β-catenin、Notch 1 mRNA表达;迁移实验观察MCF-7迁移状况。结果经免疫组化α-SMA、Vimentin、FSP抗体鉴定,证实纯化的细胞为活化的间质成纤维细胞。实验组MCF-7的Wnt1、Notch1表达明显上调,分别为对照组的2.09倍和3.75倍,而β-catenin表达下调为对照组的0.31倍;实验组MCF-7穿过膜的细胞数明显增多,为对照组的2.3倍。结论乳腺癌原代间质细胞对MCF-7 Wnt、Notch1、β-catenin mRNA表达具有调节作用,并促进MCF-7的迁移。

Objective To investigate the regulatory effects of cancer stromal cells on cancer cell lines MCF-7,and to explore the roles of microenvironment in the carcinogenesis and development of breast cancer. Methods Five specimens from patients with breast cancer were collected,and stromal cells were isolated,cultured and purified,and were identified by immunohistochemistry.Stromal cells and MCF-7 cells were co-cultured for 3 d via Transwell system（experiment group）.The expression of Wnt1,β-catenin and Notch1 mRNA in MCF-7 were detected by Real-time PCR.The effect of migration on MCF-7 by cancer stromal cells was detected.MCF-7 cultured alone with 2% FCS was served as control group. ResultsThe stromal cells purifed were fibroblasts with identification of α-SMA,vimentin and FSP antibodies by immunohistochemi-stry.The expression of Wnt1 and Notch1 in MCF-7 in experiment group was significantly upregulated,2.09 and 3.75 times higher to control group,respectively.However,the expression of β-catenin was down-regulated,0.31 times lower to control group.The number of migrating cells was significantly increased in experiment group,2.3 times larger to control group.Conclusion The breast cancer stromal cells can regulate the expression of Wnt,Notch1 and β-catenin mRNA in MCF-7,and can accelerate the migration of MCF-7.