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

作　　者：赵盛[1] 苗燕平[1] 王睿君 包丽薇[1] ZHAO Sheng;MIAO Yanping;WANG Ruijun;BAO Liwei(Department of Imaging Diagnosis,The Affiliated Hospital of Inner Mongolia Medical University,Hohhot O10050,China;Inner Mongolia Clinical Medical Research Center of Nervous System Diseases,Hohhot 010050,China)

机构地区：[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

出　　处：《中国实验诊断学》2024年第6期701-707,共7页Chinese Journal of Laboratory Diagnosis

基　　金：内蒙古自治区高等学校科学研究项目(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)。

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。Objective To explore the miR-544 in glioblastoma(glioblastoma, GBM) epithelial-mesenchymal transition(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-β group 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 

分 类 号：R739.4[医药卫生—肿瘤]