基础医学与临床 ›› 2018, Vol. 38 ›› Issue (8): 1094-1098.

• 研究论文 • 上一篇    下一篇

miR-26b对宫颈癌细胞系迁移及上皮间质转化的影响

凌燕1,彭诗维1,冯紫雯2,龚剑红1   

  1. 1. 江西省人民医院妇产科
    2. 江西省南昌县妇幼保健院 妇产科
  • 收稿日期:2017-12-08 修回日期:2018-04-29 出版日期:2018-08-05 发布日期:2018-07-24
  • 通讯作者: 龚剑红 E-mail:2274534115@qq.com
  • 基金资助:
    江西省重点研发计划项目;江西省卫生计生委科技计划

Effects of miR-26b on the migration and epithelial mesenchymal transition of cervical cancer cell line

  • Received:2017-12-08 Revised:2018-04-29 Online:2018-08-05 Published:2018-07-24

摘要: 目的 探讨miR-26b对宫颈癌细胞迁移及上皮间质转化的影响及其作用机制。方法 通过脂质体介导将miR-26b mimic转染C33A宫颈癌细胞,细胞分为阴性对照组、无义对照组和miR-26b mimic组。用实时荧光定量 PCR,检测 miR-26b及其靶基因的mRNA表达,细胞划痕实验检测细胞的迁移能力,Western blot检测E-cadherin和N-cadherin蛋白表达,用双荧光素酶基因报告载体检测 miR-26 b 靶基因的表达。结果 与正常宫颈上皮细胞相比,miR-26b mRNA在HeLa、SiHa、Caski和C33A 4种宫颈癌细胞中均低表达,其中以C33A细胞最为明显;转染miR-26b mimic后,C33A细胞中miR-26b mRNA表达水平明显上升;过表达miR-26b,可明显抑制C33A细胞的迁移能力,上调E-cadherin蛋白的表达,下调N-cadherin蛋白的表达,抑制上皮间质转化过程的发生;通过TargetScan、PicTar和miRDB软件预测显示,CXCL14、Smad4、TRAF5和EphA2可能为miR-26b潜在的靶基因。双荧光素酶实验证实miR-26 b 可直接调控Smad4的表达。结论 miR-26b 可能通过作用于Smad4的表达,调节宫颈癌细胞的迁移和上皮间质转化过程的发生。

关键词: microRNA-26b, 宫颈癌, C33A细胞, 迁移, 上皮间质转化

Abstract: Objective To investigate the effect and mechanism of miR-26b on the migration and epithelial mesenchymal transition of cervical cancer cells. Methods miR-26b mimic was transfected into C33A cervical cancer cells by liposome-mediated method. The experiment was divided into negative control group, scramble control group and miR-26b mimic group. The expression of miR-26b and its target genes mRNA were detected by real-time fluorescence quantitative PCR. The cell migration ability was examined by scratch-wound assay. The expression of E-cadherin and N-cadherin proteins were detected by Western blot. Dual luciferase gene reporter assays were used to examine whether miR-26b regulates the expression of Smad4. Results Compared with normal cervical epithelial cells, expression levels of miR-26b mRNA were all lower in SiHa, HeLa, Caski and C33A cells, among which C33A cells were the most obvious. mRNA expression levels of miR-26b were significantly increased in C33A cells by transfection of miR-26b mimic. Over-expression of miR-26b can significantly inhibit the migration of C33A cells, increase the expression of E-cadherin protein and reduce the expression of N-cadherin protein, and inhibit the process of epithelial mesenchymal transition. The potential target genes of miR-26b were predicted by TargetScan, PicTar and miRDB software. Based on the annotated functions for these genes, CXCL14, Smad4, TRAF5 and EphA2 were speculated to be the major potential target genes of miR-26b. Dual luciferase gene reporter assays confirmed that miR-26b can directly regulate the expression of Smad4. Conclusions miR-26b may inhibit the migration of cervical cancer cells and the process of epithelial mesenchymal transition by regulating the expression of Smad4.

Key words: microRNA-26b, cervical cancer, C33A cells, migration, epithelial mesenchymal transition

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