miR-544靶向TGF-β抑制神经胶质母细胞瘤细胞迁移、侵袭及EMT
MiR-544 inhibited cell migration,invasion and EMT by targted TGF-β in GBM
赵盛 1苗燕平 1王睿君 2包丽薇1
作者信息
- 1. 内蒙古医科大学附属医院影像诊断科,内蒙古呼和浩特 010050
- 2. 内蒙古自治区神经系统疾病临床医学研究中心,内蒙古呼和浩特 010050
- 折叠
摘要
目的 探究miR-544在神经胶质母细胞瘤(glioblastoma,GBM)上皮-间质转化(epithelial-mesenchymal transition,EMT)过程中的作用及机制.方法 采用RT-qPCR检测miR-544及TGF-β在GBM组织和癌旁组织、GBM转移与非转移的病人血清中的表达.在GBM细胞系U251中转染MiR-544 mimics.采用Transwell、Western boltting检测细胞迁移及侵袭,EMT相关蛋白(E-cadherin和N-cadherin)的变化.荧光素酶报告基因验证miR-544与TGF-β基因3'UTR区结合.结果 miR-544在GBM组织及转移组中表达显著降低;TGF-β显著升高(P＜0.05).荧光素酶报告证实miR-544靶向调节TGF-β3'-UTR区域.miR-544 mimics组及si-TGF-β组与NC组相比,细胞迁移与侵袭数目减少(P＜0.05);E-cadherin蛋白表达增加,N-cadherin蛋白表达减少(P＜0.05).miR-544mimics+pcDNA-TGF-β组部分逆转了 miR-544 mimics对细胞迁移与侵袭和EMT相关蛋白的表达的作用(P＜0.05).结论 miR-544靶向调控TGF-β是其参与EMT过程的作用机制之一.
Abstract
Objective To explore the miR-544 in glioblastoma(glioblastoma,GBM)epithelial-mesenchymal transi-tion(EMT).Methods RT-qPCR was used to detect the expression of miR-544 and TGF-β in GBM tissues and adjacent tissues,and serum of GBM patients with and without metastasis.MiR-544 mimics were transfected into GBM cell line U251.Transwell and Western blotting were used to detect the changes of cell migration and invasion,and EMT-related proteins(E-cadherin and N-cadherin).Luciferase reporter gene verified the binding of miR-544 to the 3'utr region of TGF-β gene.Results The expression of miR-544 was significantly decreased in GBM tissues and metastasis groups.TGF-β significantly increased(P＜0.05).Luciferase reporter assay confirmed that miR-544 targeted the 3'-UTR region of TGF-β.Compared with NC group,the number of cell migration and invasion in miR-544 mimics group and si-TGF-βgroup was decreased(P＜0.05);The expression of E-cadherin protein was increased,while the expression of N-cadherin protein was decreased(P＜0.05).The miR-544mimics+pcDNA-TGF-β group partially reversed the effect of miR-544mimics on the expression of cell migration,invasion and EMT related proteins(P＜0.05).Conclusion miR-544 is involved in EMT by targeting TGF-β.
关键词
miR-544/EMT/TGF-β/胶质母细胞瘤Key words
miR-544/EMT/TGF-β/glioblastoma multiforme引用本文复制引用
基金项目
内蒙古自治区高等学校科研项目(NJSY23081)
内蒙古医科大学联合项目(YKD2023LH061)
内蒙古自治区自然科学基金(2017MS0833)
内蒙古自治区科技计划(2019GG9044)
出版年
2024
NETL
NSTL
万方数据