微小 RNA-744-5p靶向整合素连接激酶对膀胱癌细胞增殖、迁移、侵袭的影响

目的 探讨微小RNA-744-5p(miR-744-5p)对膀胱癌细胞增殖、迁移和侵袭的影响及可能机制.方法 选取2016年2月至2018年11月枣庄矿业集团枣庄医院接收的经病理确诊的56例膀胱癌病人病理组织切片,采用实时荧光定量逆转录聚合酶链反应(qRT-PCR)检测56例膀胱癌组织和对应癌旁组织中miR-744-5p表达水平.分别转染miR-744-5p模拟物(mimics)、miR-744-5p抑制剂或共转染miR-744-5p mimics与整合素连接激酶(ILK)过表达载体至膀胱癌BIU-87、T739细胞,MTT检测细胞增殖,Transwell检测细胞迁移和侵袭,蛋白质印迹法(Western Blotting)检测过细胞中细胞周期蛋白(Cyclin D1)、P21、基质金属蛋白酶(MMP-2)和钙黏附蛋白E(E-cadherin)蛋白表达.双荧光素酶报告基因实验验证miR-744-5p与ILK调控关系.结果 与癌旁组织相比,膀胱癌组织中miR-744-5p表达水平显著降低(0.98±0.10)比(0.25±0.02),P<0.05].过表达miR-744-5p后,BIU-87和T739细胞OD值、迁移数和侵袭数及Cyclin D1和MMP-2蛋白表达降低(P<0.05),而P21和E-cadherin蛋白表达升高(P<0.05).抑制miR-744-5p后,BIU-87和T739细胞OD值、迁移数和侵袭数升高(P<0.05).miR-744-5p靶向结合并负调控ILK,过表达ILK逆转过表达miR-744-5p对BIU-87和T739细胞增殖、迁移和侵袭的影响.结论 膀胱癌组织中miR-744-5p呈低表达,过表达miR-744-5p可阻碍膀胱癌细胞增殖、迁移和侵袭,而抑制miR-744-5p则起相反的作用,其可能通过靶向负调控ILK表达发挥作用.

Effect of miR-744-5p on proliferation, migration and invasion of bladder cancer cells and its mechanism by targeting ILK

Objective To investigate the effect of miR-744-5p on proliferation, migration and invasion of bladder cancer BIU-87 cells and its mechanism.Methods A total of 56 pathologically confirmed patients with bladder cancer in Zaozhuang Mining Group Zaozhuang Hospital from February 2016 to November 2018 were selected. RT-qPCR was used to detect the expression of miR-744-5p in 56 bladder cancer tissues and corresponding adjacent tissues. After miR-744-5p mimics or miR-744-5p inhibitor was transfected into bladder cancer BIU-87 and T739 cells, or miR-744-5p mimics and integrin-linked kinase (ILK) overexpression vectors were co-transfected into bladder cancer BIU-87 and T739 cells, and then MTT was used to detect the proliferation. Transwell was used to detect themigration and invasion, and Western Blot was used to detect the protein levels of Cyclin D1, P21, MMP-2 and E-cadherin. The dual luciferase reporter gene assay validated the relationship between miR-744-5p and ILK.Results The expression level of miR-744-5p in bladder cancer tissues was significantly lower than that in adjacent tissues (0.98±0.10) vs. (0.25±0.02), P<0.05]. After overexpression of miR-744-5p, the OD value, the number of migration and invasion and the protein levels of Cyclin D1 and MMP-2 protein in BIU-87 and T739 cells were decreased (P<0.05), but the protein levels of P21 and E-cadherin were increased (P<0.05). After inhibiting miR744-5p, the OD value, the number of migration and invasion of BIU-87 and T739 cells were increased (P<0.05). miR-744-5p targeted and negatively regulated ILK. Overexpression of ILK reversed the effects of overexpression of miR-744-5p on the proliferation, migration and invasion of BIU-87 and T739 cells.Conclusion The expression of miR-744-5p in bladder cancer tissues is low. Overexpression of miR-744-5p can hinder the proliferation, migration and invasion of bladder cancer cells, but inhibition of miR-744-5p has the opposite effect, which may play a role by targeting and negatively regulating expression of ILK .