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吴悠, 汤婷婷, 朱琴华, 金亮, 潘怡. miR-29a促进人乳腺癌MCF-7细胞体外迁移和侵袭的机制[J]. 中国药科大学学报, 2018, 49(3): 348-353. DOI: 10.11665/j.issn.1000-5048.20180314
引用本文: 吴悠, 汤婷婷, 朱琴华, 金亮, 潘怡. miR-29a促进人乳腺癌MCF-7细胞体外迁移和侵袭的机制[J]. 中国药科大学学报, 2018, 49(3): 348-353. DOI: 10.11665/j.issn.1000-5048.20180314
WU You, TANG Tingting, ZHU Qinhua, JIN Liang, PAN Yi. Mechanisms of miR-29a in migration and invasion of breast cancer MCF-7 cells in vitro[J]. Journal of China Pharmaceutical University, 2018, 49(3): 348-353. DOI: 10.11665/j.issn.1000-5048.20180314
Citation: WU You, TANG Tingting, ZHU Qinhua, JIN Liang, PAN Yi. Mechanisms of miR-29a in migration and invasion of breast cancer MCF-7 cells in vitro[J]. Journal of China Pharmaceutical University, 2018, 49(3): 348-353. DOI: 10.11665/j.issn.1000-5048.20180314

miR-29a促进人乳腺癌MCF-7细胞体外迁移和侵袭的机制

Mechanisms of miR-29a in migration and invasion of breast cancer MCF-7 cells in vitro

  • 摘要: 探讨微小核酸miRNA-29a对人乳腺癌细胞MCF-7体外迁移和侵袭的促进作用及其机制。采用miR-29a模拟物及miR-29a抑制剂分别上调和下调miR-29a的表达;采用细胞划痕法和Transwell小室法检测miR-29a对细胞迁移和侵袭能力的影响;采用Targetscan7.1数据库预测miR-29a的靶基因;荧光素酶报告法验证miR-29a的靶基因;Western blot和实时荧光定量PCR验证其表达结果。结果表明:miR-29a可显著促进MCF-7细胞的迁移和侵袭能力。Targetscan软件预测HBP1 可能为miR-29a的靶基因,荧光素酶报告实验显示miR-29a靶向HBP1 基因的3′-UTR区。Western blot和实时荧光定量PCR结果显示,miR-29a下调了HBP1蛋白水平的表达,而mRNA水平则无明显变化。研究结果揭示在乳腺癌中高表达的miR-29a通过下调HBP1,从而使乳腺癌细胞获得高的迁移和侵袭能力,促进乳腺癌的转移。

     

    Abstract: The aim of this study was to investigate the effect and mechanisms of miR-29a in migration and invasion of human breast cancer MCF-7 cells in vitro. MCF-7 cells were treated with miR-29a mimic or miR-29a inhibitor to up-regulate/down-regulate the expression level of miR-29a. Wound-healing assay and transwell chamber were employed to determine cell migration and invasion in vitro. The target gene of miR-29a was predicted with the Targetscan7. 1 database and verified through luciferase reporter method. The effects of miR-29a on the expression of the potential target were detected by Western blot and real-time PCR. Results showed that in vitro migration and invasion ability of MCF-7 cells was increased significantly by miR-29a, which could target HBP1 in the 3′-UTR region. The protein expression of HBP1 was decreased by miR-29a overexpression. However, the alteration of miR-29a had no significant effect on the expression of HBP1 mRNA. The results validated that miR-29a, highly expressed in breast cancer, could down-regulate HBP1, which in turn promotes migration and invasion ability of breast cancer cells, thus promoting breast cancer metastasis.

     

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