miR －181c抑制胶质母细胞瘤细胞株 T98G侵袭能力的机制

Mechanism of miR-181c inhibiting the invasion of glioblastoma cell line T 98G

目的：探讨miRNA －181c（miR－181c）抑制胶质母细胞瘤（GBM ）细胞株T98G侵袭的机制。方法应用实时定量PCR、蛋白质印迹法（Western Blot）检测miR－181c与转化生长因子β（TGF－β）相关基因的表达水平，应用Transwell实验检测miR－181c与TGF－β对胶质瘤细胞侵袭的影响，利用分子克隆技术设计、克隆目的基因的3＇端非翻译区（3＇－UTR）及其突变区域，通过荧光素酶分析miR－181c与目的基因的直接结合情况。结果 miR－181c转染后与TGF－β通路相关基因转化生长因子β受体1（TGFBR1）、TGFBR2和转化生长因子β受体相关蛋白1（TGFBRAP1）的mRNA及蛋白表达水平下降，差异有统计学意义（P ＜0．05），miR－181c通过影响TGF－β信号减弱T98G细胞的侵袭性，且TGFBR1、TGFBR2和TGFBRAP1的3＇－UTR是miR －181c的直接作用靶点。结论 miR－181c可能通过调控 TGF －β信号通路抑制胶质瘤细胞株 T98G的侵袭能力，可以做为GBM 治疗的重要分子。

Objective To investigate the mechanism of miR -181c inhibiting the invasion of glio‐blastoma cell line T98G .Methods Using RT -PCR and Western Blot to detect the expression of miR -181c and TGF-βrelated genes .The effect of miR -181c and TGF -βon the invasion of glioma cells was measured by Transwell .Using molecular cloning technology to design and clone the target gene 3＆#39;-UTR and its mutation region .Using luciferase assay to analyse the combination of miR -181c and target genes .Results mRNA and protein expression of miR -181c and TGF -βpathway associated genes TG‐FBR1 ,TGFBR2 and TGFBRAP1 decreased （P〈0 .05） .miR-181c could weaken the invasion of T98G cells by influencing TGF -βsignal and the 3＆#39;-UTR of TGFBR1 ,TGFBR2 and TGFBRAP1 were direct targets of miR-181c .Conclusions miR -181c can inhibit the invasive ability of glioma cell line T 98G by regulating the TGF -βsignaling pathway ,and it can be an important molecule for glioblastoma treatment .