上海交通大学学报（医学版）

›› 2012, Vol. 32 ›› Issue (6): 699-.doi: 10.3969/j.issn.1674-8115.2012.06.003

• 论著（基础研究） • 上一篇    下一篇

miR-506通过靶向Slug调节上皮间充质转化对乳腺癌细胞侵袭和转移的抑制作用

梁永俊1, 周佽想2, 宇小婷2, 赵 倩2   

  1. 1.上海交通大学医学院/中国科学院上海生命科学研究院健康科学研究所, 2.上海交通大学基础医学院病理生理学教研室, 上海 200025
  • 出版日期:2012-06-28 发布日期:2012-07-02
  • 通讯作者: 赵 倩, 电子信箱: qzhao@shsmu.edu.cn。
  • 作者简介:梁永俊（1986—）, 男, 硕士生；电子信箱: liangyj0214@sina.com。
  • 基金资助:

    国家自然科学基金（81172521, 81170505）

miR-506 targets Slug to regulate epithelial-to-mesenchymal transition and metastasis of breast cancer

LIANG Yong-jun1, ZHOU Ci-xiang2, YU Xiao-ting2, ZHAO Qian2   

  1. 1.Institute of Health Sciences, Shanghai Jiaotong University School of Medicine &|Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200025, China；2.Department of Pathophysiology, Basic Medical College, Shanghai Jiaotong University, Shanghai 200025, China
  • Online:2012-06-28 Published:2012-07-02
  • Supported by:

    National Natural Science Foundation of China, 81172521, 81170505

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摘要：

目的 探讨miR-506通过靶向Slug调节上皮间充质转化对乳腺癌细胞侵袭和转移的抑制作用。方法 采用实时荧光定量PCR检测8株乳腺癌细胞系（MCF7、BT474、SK-BR-3、MDA-MB-453、MDA-MB-468、HCC1937、BT549和MDA-MB-231）中miR-506的表达量，以正常乳腺上皮细胞MCF10A为对照。通过脂质体转染的方法在MDA-MB-231和BT549细胞系中瞬时过表达miR-506模拟物（设阴性对照），通过划痕实验、Transwell迁移和侵袭实验观察细胞运动能力的变化；观察MDA-MB-231和BT549细胞系转染miR-506前后细胞形态学变化；Western blotting法检测E-cadherin、Vimentin和Slug蛋白表达的变化。荧光素酶报告基因分析实验检测MDA-MB-231细胞中miR-506过表达对野生型荧光素酶活性的影响。结果 miR-506在各乳腺癌细胞中的表达量相对于正常乳腺上皮细胞MCF10A显著降低（P<0.01）。miR-506过表达显著抑制MDA-MB-231和BT549细胞的迁移和侵袭能力，与阴性对照的差异有统计学意义（P<0.01）。MDA-MB-231和BT549细胞瞬时转染miR-506 48 h后发生明显的形态学改变；Western blotting结果显示miR-506过表达在MDA-MB-231和BT549细胞中均上调E-cadherin的表达，下调Vimentin的表达。Western blotting检测结果显示MDA-MB-231细胞转染miR-506后Slug蛋白水平明显低于阴性对照；荧光素酶报告基因分析结果显示miR-506过表达显著抑制野生型荧光素酶的活性。结论 在乳腺癌中，miR-506可以通过靶向转录因子Slug诱导细胞发生上皮间充质转化，进而抑制细胞的侵袭和转移能力，这是miR-506参与乳腺癌发生和发展的可能途径之一。

关键词: miR-506, 乳腺癌, 上皮间充质转化, Slug

Abstract:

Objective To investigate the role of miR-506 targeting Slug in regulation of epithelial-to-mesenchymal transition and metastasis of breast cancer. Methods The expression of miR-506 in eight breast cancer cell lines (MCF7, BT474, SK-BR-3, MDA-MB-453, MDA-MB-468, HCC1937, BT549 and MDA-MB-231) was determined by Real-Time fluorescence quantitative PCR, and human normal mammary epithelial cell MCF10A was served as control. miR-506 mimics were transiently overexpressed in MDA-MB-231 and BT549 cells through liposome transfection(negative control). The mobility of cells was detected by wound healing assay and Transwell migration and invasion assay. The morphological change of cells after transfection with miR-506 was observed, and the expression of E-cadherin, Vimentin and Slug protein was determined by Western blotting. The effect of miR-506 overexpression in MDA-MB-231 cells on wild type luciferase was determined by luciferase reporter assay. Results The expression of miR-506 in breast cancer cells was significantly lower than that in MCF10A cells (P<0.01). The overexpression of miR-506 significantly inhibited the migration and invasion ability of MDA-MB-231 and BT549 cells, which was significantly different from that of negative control (P<0.01). Obvious morphological change was observed in MDA-MB-231 and BT549 cells 48 h after transient transfection with miR-506. Western blotting indicated that miR-506 overexpression increased the expression of E-cadherin and decreased the expression of Vimentin in MDA-MB-231 and BT549 cells. Western blotting revealed that the expression of Slug protein in MDA-MB-231 cells after transfection with miR-506 was significantly lower than that in negative control, and luciferase reporter assay demonstrated that miR-506 overexpression significantly inhibited the activity of wild type luciferase. Conclusion MiR-506 targets Slug to regulate epithelial-to-mesenchymal transition and inhibit metastasis of breast cancer, which may be one of the ways for miR-506 to participate in the development of breast cancer.

Key words: miR-506, breast cancer, epithelial-mesenchymal transition, Slug