基础医学与临床 ›› 2018, Vol. 38 ›› Issue (9): 1244-1250.

• 研究论文 • 上一篇    下一篇

人骨髓间充质干细胞促进乳腺癌MCF7细胞上皮间质转化

许峰,金晓霞,施民新   

  1. 南通市肿瘤医院
  • 收稿日期:2017-05-26 修回日期:2017-10-01 出版日期:2018-09-05 发布日期:2018-09-10
  • 通讯作者: 施民新 E-mail:smx654321@126.com
  • 基金资助:
    南通市卫生局青年科研基金项目

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

  • Received:2017-05-26 Revised:2017-10-01 Online:2018-09-05 Published:2018-09-10

摘要: 目的 探讨在人骨髓间充质干细胞(hBMSCs)与乳腺癌细胞MCF-7共培养后,MCF-7细胞上皮间质转化能力是否受到影响,并检测共培养条件下上皮间质转换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hBMSCs,利用Transwell小室将hBMSCs与乳腺癌细胞系MCF-7共培养,光镜观察MCF-7细胞的形态结构,是否发生上皮间质转换的特征性改变。利用实时定量PCR、Western blot、免疫荧光检测上皮间质转换标志蛋白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细胞上皮间质转换的分子标志物的水平;CCK8检测MCF-7细胞的增殖活力;流式细胞术检测增殖周期的变化;transwell检测MCF-7细胞侵袭及转移能力。结果 与对照组相比,与hBMSCs共培养之后的MCF-7细胞表现出明显增强的上皮间质转换能力(P<0.05)。培养体系中的条件培养液刺激MCF-7细胞,其上皮间质转换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发生上皮间质转换。骨髓间充质干细胞与乳腺癌细胞可以通过旁分泌信号促进乳腺癌上皮间质转换,进一步增强其增殖及转移能力。

关键词: 关键词:人骨髓间充质干细胞, 乳腺癌, 上皮间质转化, 旁分泌机制

Abstract: Objective To explore whether epithelial-mesenchymal transition(EMT) is affected after the human bone marrow mesenchymal stem cells (hBMSCs) co-cultured with MCF-7 breast cancer cells. Study the effect of paracrine factors on proliferation and migration in MCF7 cells. Methods hBMSCs was isolated successfully in vitro.Co-cultured hBMSCs with breast cancer cell line MCF-7 by transwell chamber, then observed the morphological changes of MCF-7 cells. The expression levels of E-cadherin, vimentin,α-SMA and β-catenin were detected by real-time quantitative PCR,Western blot and immunofluorescence. Collected conditional-culture medium from co-culture system and 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 hBMSCs,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). Conclusion hBMSCs could induce EMT of breast cancer cells and promote the prolifertion,migration and invasion capacity of breast cancer cells by paracrine signal.

Key words: Key words: human bone marrow mesenchymal stem cells, Breast cancer, Epithelial-mesenchymal transition, paracrine mechanism

中图分类号: