miR-193b通过负向调控MCT7基因的表达抑制人胶质瘤U251细胞的迁移与侵袭

发布时间：2018-03-21 00:42

本文选题：胶质瘤　切入点：miR-b　出处：《厦门大学学报(自然科学版)》2017年05期 　论文类型：期刊论文

【摘要】：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.
[Abstract]:In order to investigate the effect of miR-193b on the migration and invasion of human glioma U251 cells, transient transfection of miR-193b mimicswith HiPerFect transfection reagent was used to up-regulate the expression of miR-193b in U251 cells. The transfection efficiency was detected by real-time fluorescence quantitative PCR qRT-PCR. The effects of miR-193b on the migration and invasion of U251 cells were examined by cell scratch assay and invasion assay. The results showed that overexpression of miR-193b could inhibit the migration and invasion of U251 cells. Bioinformatics software was used to predict the effect of miR-193b on the migration and invasion of U251 cells. It is suggested that monocarboxylate transporter 7hmCT7) gene may be a potential target gene of miR-193b. Furthermore, the construction of double luciferase reporter gene system at the transcriptional level proves that MCT7 is the target gene regulated by miR-193b, qRT-PCR and Western blot of protein imprinting are found in mRNA respectively. It is further demonstrated that the expression of MCT7 gene is negatively regulated by miR-193b, which suggests that the expression of MCT7 gene may inhibit the migration and invasion of U251 cells by negatively regulating the expression of MCT7 gene.
【作者单位】：厦门大学附属第一医院神经外科;
【基金】：厦门市科技惠民计划项目(2013S0001)
【分类号】：R739.41

【相似文献】