miR-34a通过MMP2抑制脑胶质瘤细胞侵袭和转移  

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

作　　者：涂其武 沈晓黎[1] TU Qi-wu;SHEN Xiao-li(Second Affiliated Hospital of Nanchang University,Nanchang 33000)

机构地区：[1]南昌大学第二附属医院,江西南昌330000

出　　处：《基础医学与临床》2018年第8期1115-1120,共6页Basic and Clinical Medicine

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和转移的影响。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-q PCR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细胞侵袭和迁移能力。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抑制细胞侵袭和迁移(P<0.05)。结论在胶质瘤细胞中miR-34a下调MMP2,进而抑制细胞侵袭和迁移能力。Objective To investigate the effect of miR-34 a on the invasion and metastasis of glioma cells by regulating the expression of matrix metalloproteinase 2（ MMP2） as a target gene of miR-34 a. Methods U87 and A172 with MMP2 high expression were selected to transfect miR-34 a mimics and miR-34 a inhibitors. To observe the expression of miR-34 a and MMP2 in the multiple glioma cell lines by RT-q PCR and Western blot. After predicting the region of miR-34 a and MMP2 with microRNA database,construct MMP2 ＇ UTR wild-type and mutant luciferase plasmids and miR-34 a plasmids,to observe the wild type and mutant luciferase activity changes by the luciferase reporter gene experiments. The invasion and migration of U87 in blank control group,MMP2 group and miR-34 a/MMP2 group were measured by Transwell and wound healing assay. Results MMP2 protein and mRNA level was decreased after over-expression of miR-34 a,and increased after decreasing miR-34 a as compared with the normal group（ P〈0. 05）; miR-34 a combined with MMP2 mRNA 3＇UTR region reduce MMP2 the level of mRNA,miR-34 a inhibited cell migration and invasion by down-regulating MMP2.Conclusions miR-34 a can down-regulate the invasion and migration of MMP2 in glioma cells.

关 键 词：MIR-34A 基质金属蛋白酶2(MMP2) 胶质瘤 侵袭和转移 

分 类 号：R739.41[医药卫生—肿瘤]