miR-186通过负调控Smad6对胶质瘤细胞迁移和上皮间质转化的影响  

miR-186 inhibits the migration and epithelial-mesenchymal transition of human glioma cell lines by targeting Smad6

作　　者：刘艳[1] 许路[1] 邵阿末[1] 曾金艳[1] 杨国青[2] LIU Yan;XU Lu;SHAO Amo;ZENG Jinyan;YANG Guoqing(Wuxi Higher Health Vocational Technology School,Wuxi 214028,China;Department of Pathology,Xi’an Central Hospital,Xi’an 710003)

机构地区：[1]无锡卫生高等职业技术学校,江苏无锡214028 [2]西安市中心医院病理科,西安710003

出　　处：《中国比较医学杂志》2020年第9期25-31,共7页Chinese Journal of Comparative Medicine

基　　金：江苏省无锡市卫生计生委科研面上项目(MS201765)。

摘　　要：目的研究miR-186通过负调控Smad6对人胶质瘤细胞迁移和上皮间质转化的影响。方法qRTPCR检测星形胶质细胞及胶质瘤细胞中miR-186和Smad6的表达,Western blot检测Smad6蛋白的表达水平;细胞分四组:空白对照组(U251)、模拟物组(miR-186 mimics)、质粒组(pc-Smad6)、共转染组(miR-186 mimics+pc-Smad6)。Western blot检测各组细胞Smad6、上皮间质转化相关蛋白E-cadherin和Vimentin的表达水平;划痕实验检测各组细胞的迁移情况。结果与正常星形胶质细胞比,人胶质瘤细胞系U251中miR-186表达明显降低,而Smad6表达明显升高(P<0.05)。较对照组,Smad6蛋白在miR-186 mimics组细胞中的表达显著下降,而在pc-Smad6组细胞中的表达则明显增加(P<0.05);miR-186 mimics组细胞迁移能力显著降低,而pc-Smad6组明显增强(P<0.05);miR-186 mimics组E-cadherin蛋白的表达显著升高,而Vimentin蛋白表达明显降低(P<0.05),在pc-Smad6组中两种分子的表达情况相反(P<0.05)。对比pc-Smad6组,miR-186 mimics+pc-Smad6组Smad6蛋白的表达水平明显降低(P<0.05);miR-186 mimics+pc-Smad6组细胞的迁移能力明显降低(P<0.05);miR-186 mimics+pc-Smad6组Vimentin表达明显降低,而E-cadherin的表达显著升高(P<0.05)。结论miR-186可通过调控Smad6的表达而抑制人胶质瘤细胞迁移和上皮间质转化。Objective We investigated the effect of mothers against decapentaplegic homolog 6(Smad6)on the migration and epithelial-mesenchymal transition of human glioma cell lines by targeting microRNA-186(miR-186).Methods The expression of miR-186 and Smad6 at the mRNA and protein level(for Smad6)was determined by RTqPCR and/or Western blot in astrocytes and glioma cells.The cells were divided into four groups:a blank control group(U251),a mimic group(miR-186 mimics),a plasmid group(pc-smad6),and a co-transfection group(miR-186 mimics+pc-smad6).The expression of Smad6,E-cadherin,and Vimentin at the mRNA and protein level was determined by RTqPCR and Western blot in each group.Migration ability was measured using a wound healing experiment.Results Compared with astrocytes,the expression of miR-186 in human glioma cell line U251 decreased significantly,while the expression of Smad6 mRNA increased significantly(P<0.05).Compared with the control group,the expression level of the Smad6 protein in the miR-186 mimic group decreased significantly,while the level of the Smad6 protein in the pcsmad6 group increased significantly(P<0.05).Compared with the pc-smad6 group,the expression level of the Smad6 protein in the miR-186 mimics+pc-smad6 group significantly decreased(P<0.05).Compared with the control group,the migration ability of the miR-186 mimics group decreased significantly,while the pc-smad6 group’s migration ability was enhanced.Compared with the pc-smad6 group,the migration ability of the miR-186 mimics+pc-Smad6 group decreased significantly(P<0.05).Compared with the control group,E-cadherin expression in the miR-186 mimics group increased significantly,while Vimentin protein expression decreased significantly(P<0.05).The expression of the two molecules in the pc-smad6 group was the opposite(P<0.05).Compared with the pc-smad6 group,vimentin expression in the miR-186 mimics+pc-smad6 group decreased significantly,while E-cadherin expression increased significantly(P<0.05).Conclusions miR-186 may regulate migration and ep

关 键 词：人胶质瘤 miR-186 SMAD6 迁移 上皮间质转化 

分 类 号：R-33[医药卫生]