人骨髓间充质干细胞促进乳腺癌细胞系MCF7上皮间质转化

Human bone marrow mesenchymal stem cells induce epithelial-mesenchymal transition in breast cancer cells line MCF-7

目的探讨在人骨髓间充质干细胞(h BMSCs)与乳腺癌细胞系MCF-7共培养后,MCF-7细胞上皮间质转化(MET)能力是否受到影响,并检测共培养条件下上皮间质转化是否是通过旁分泌因素影响MCF-7细胞的增殖及迁移能力的。方法体外成功分离h BMSCs,利用Transwell小室共培养h BMSCs与乳腺癌细胞系MCF-7,光镜观察MCF-7细胞的形态结构,是否发生上皮间质转化的特征性改变。利用实时定量PCR、Western blot、免疫荧光检测上皮间质转化标志蛋白E-cadherin、vimentin、α-SMA及β-catenin的表达。收集共培养体系中的条件培养液,作用MCF-7细胞,Western blot鉴定MCF-7细胞上皮间质转化的分子标志物的水平;CCK8法检测MCF-7细胞的增殖活力;流式细胞计量术检测细胞增殖周期的变化;Transwell小室法检测MCF-7细胞侵袭及转移能力。结果与对照组相比,与h BMSCs共培养之后的MCF-7细胞表现出明显增强的上皮间质转化能力(P<0.05)。培养体系中的条件培养液刺激MCF-7细胞,其上皮间质转化特征及增殖和迁移能力增强(P<0.01)。结论人骨髓间充质干细胞可诱导乳腺癌细胞系发生上皮间质转化。骨髓间充质干细胞与乳腺癌细胞系可以通过旁分泌信号促进乳腺癌上皮间质转化,进一步增强其增殖及转移能力。

Objective To explore whether epithelial-mesenchymal transition（ EMT） is affected after the human bone marrow mesenchymal stem cells（ h BMSCs） co-cultured with MCF-7 breast cancer cells line. Study the effect of paracrine factors on proliferation and migration in MCF7 cells. Methods h BMSCs were successfully isolated in vitro.Co-cultured h BMSCs with breast cancer cell line MCF-7 by transwell chamber,then observed the morphological changes of MCF-7 cells. The expression of E-cadherin,vimentin,α-SMA and β-catenin was detected by realtime quantitative PCR,Western blot and immunofluorescence. Collected conditional-culture medium from co-culture system acted on MCF7 cells; Western blot analysis tested EMT markers of MCF7 cells; CCK8 assay deteced cell growth activity; flow cytometry tested cell cycle; Transwell investigated invasion and metastasis of MCF7 cells.Results After co-cultured with h BMSCs,MCF7 cells showed incresed EMT ability（ P〈0. 05）. Stimulated with conditional-medium from the co-cultured system,the EMT characteristics and proliferation,migration ability of MCF7 cells were enhanced（ P〈0. 01）. Conclusions h BMSCs could induce EMT of breast cancer cells line and promote the prolifertion,migration and invasion capacity of breast cancer cells line by paracrine signal.