为探讨miR-193b对人胶质瘤U251细胞的迁移和侵袭能力的影响,利用HiPerFect转染试剂瞬时转染miR-193b模拟物(mimics)上调U251细胞中miR-193b的表达,通过实时荧光定量PCR(qRT-PCR)检测其转染效率,然后进行细胞划痕实验及侵袭实验,分别检测miR-193b对U251细胞迁移及侵袭能力的影响.结果显示过表达miR-193b可抑制U251细胞迁移和侵袭的能力.应用生物信息学软件预测,提示单羧酸转运蛋白7(MCT7)基因可能是miR-193b的潜在靶基因.进而构建双荧光素酶报告基因系统从转录水平上证明MCT7为miR-193b调控的靶基因; qRT-PCR及蛋白质免疫印记(Western blot)分别在mRNA和蛋白水平上进一步证明MCT7基因的表达受miR-193b的负调控.由此可见,miR-193b很可能通过负向调控MCT7基因的表达来抑制U251细胞的迁移与侵袭.
To investigate the effect of miR-193b on the migration and invasion of human glioma cell line U251,upregulated expression level of miR-193b in glioma U251 cells was achieved through transfecting with miR-193b mimics using HiPerFect transfection reagent.The transfection efficiency of miR-193b was detected using qRT-PCR,and cell scratch assay and invasive experiments were used to detect the abilities of miR-193b on migration and invasion of glioma cells.Results showed that the over-expression of miR-193b can inhibit the ability of U251 cells in migration and invasion.Meanwhile,bioinformatic software was used to predict the potential miR-193b target genes,and we found that MCT7 was the potential target gene of miR-193b.Subsequently,the dual luciferase report gene system,qRT-PCR and Western blot all revealed that miR-193b negatively regulated the expression of MCT7 gene.It is suggested that miR-193b propably inhibits the migration and invasion of U251 glioma cells by negatively regulating the expression of MCT7 gene.
