基础医学与临床 ›› 2018, Vol. 38 ›› Issue (8): 1115-1120.

• 研究论文 • 上一篇    下一篇

miR-34a通过MMP2抑制脑胶质瘤细胞侵袭和转移

涂其武1,沈晓黎2   

  1. 1. 江西省中医药大学洪都中医院
    2. 南昌大学附属第二医院
  • 收稿日期:2017-03-06 修回日期:2017-12-04 出版日期:2018-08-05 发布日期:2018-07-24
  • 通讯作者: 沈晓黎 E-mail:shen104xiao@163.com

miR-34a inhibits the invasion and metastasis of glioma cells through MMP2

  • Received:2017-03-06 Revised:2017-12-04 Online:2018-08-05 Published:2018-07-24
  • Contact: Xiao liSHEN E-mail:shen104xiao@163.com

摘要: 目的 探讨miR-34a通过其靶基因基质金属蛋白酶2(MMP2)对脑胶质瘤细胞侵袭转移的影响。方法 选取U87和A172脑胶质瘤细胞,分组转染miR-34a 摸拟物和抑制剂;利用RT-qPCR及Western blot观察脑胶质瘤细胞中miR-34a和MMP2的表达;经过microRNA数据库预测miR-34a与MMP2结合区域;构建MMP2 3’UTR野生型与突变型的荧光素酶质粒和miR-34a质粒,进行荧光素酶报告基因实验;Transwell侵袭和划痕实验观察U87细胞侵袭迁移能力。结果 经筛选选用MMP2高表达细胞株U87和A172,过表达miR-34a后发现MMP2表达水平降低(p<0.05),降低miR-34a则MMP2表达升高(p<0.05);miR-34a结合MMP2 mRNA 3’UTR的特定序列降低MMP2水平(p<0.05);miR-34a通过MMP2抑制细胞侵袭迁移(p<0.05)。结论 在胶质瘤细胞中miR-34a下调MMP2进而抑制细胞侵袭迁移能力。

关键词: 关键词:miR-34a, 基质金属蛋白酶2(MMP2), 胶质瘤, 侵袭迁移

Abstract: Objective To investigate the effect of miR-34a on the invasion and metastasis of glioma cells by regulating the expression of matrix metalloproteinase 2 (MMP2), which is a target gene of miR-34a.Methods Compared with human normal brain glial cells (HEB),U87 and A172 with MMP2 high expression were selected to transfect miR-34a mimics and miR-34a inhibitors.To observe the expression of miR-34a and MMP2 in the multiple glioma cell lines by RT-qPCR and western blot.After predicting the regional of miR-34a and MMP2 with microRNA database, construct MMP2 3'UTR wild-type and mutant luciferase plasmids and miR-34a plasmids, to observe the wild type and mutant luciferase activity changes by the luciferase reporter gene experiments. The invasion and migration abilities of U87 in blank control group, MMP2 group and miR-34a /MMP2 group were measured by Transwell and wound healing assay. Results To select U87 and A172 , MMP2 protein and mRNA levels were decreased after over-expression of miR-34a, and were increased after decreasing miR-34a,compared with the normal group, the p value of protein and mRNA was less than 0.05; miR-34a combined with MMP2 mRNA 3 'UTR region to reduce the MMP2 the level of mRNA, miR-34a inhibited cell migration and invasion by down-regulating MMP2.Conclusion miR-34a can down-regulate the invasion and migration ability of MMP2 in glioma cells.

Key words: Keywords: miR-34a, matrix metalloproteinase 2(MMP2), glioma, invasion and migration

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