miRNA-491调节胶质母细胞瘤细胞凋亡和上皮间质转化的研究  

The miRNA-491 regulates apoptosis and epithelial-mesenchymal transition in glioblastoma cells

作　　者：郭小叶[1] 王佳[1] 冒平[1] 李奇[1] 廉海平[1] 白晓斌[1] 宋锦宁[1] GUO Xiaoye;WANG Jia;MAO Ping;LI Qi;LIAN Haiping;BAI Xiaobin;SONG Jinning(Department of Neurosurgery,First Affiliated Hospital of Xi’an Jiaotong University,Xi’an 710061,China)

机构地区：[1]西安交通大学第一附属医院神经外科

出　　处：《山西医科大学学报》2019年第10期1329-1336,共8页Journal of Shanxi Medical University

基　　金：国家自然科学基金青年项目(81602207,81802502);陕西省社会发展领域一般项目(2019SF-152)

摘　　要：目的研究miRNA-491(miR-491)对胶质母细胞瘤细胞上皮间质转化和凋亡的调控作用和机制。方法收集新鲜胶质瘤(glioblastoma multiforme,GBM)13例和对应的癌旁组织(nontumorous tissues,NT)13例。体外培养神经胶质细胞(normal human astrocytes,NHA)和神经胶质母细胞瘤细胞U-87 MG和U-118 MG。将U-118MG细胞分为无处理组(control)、miR-491的拟似物转染组(mimic)、mimic阴性对照组(mimic-NC)和mimic联合表达αB-crystallin组(mimic+αB-crystallin)。CCK-8法评估细胞增殖,RT-qPCR和或Western blot检测miR-491、αB-crystallin、Twist1、E-cadherin、N-cadherin、Bax、Bcl-2的表达。荧光素酶报告基因验证miR-491和αB-crystallin基因3′UTR区的结合。结果GBM组织与NT组织比较,U-87 MG和U-118 MG分别与NHA比较,miR-491显著降低(P<0.05)。与control组比较,mimic组αB-crystallin、N-cadherin、Twist1的表达水平下调,细胞凋亡率上调(P<0.05),E-cadherin的水平上调(P<0.05)。荧光素酶报告结果证实miR-491与αB-crystallin基因3′UTR区的结合。mimic+αB-crystallin组部分逆转mimic对于凋亡标记物和上皮间质转化标记物的表达(P<0.05)。结论miR-491抑制胶质母细胞瘤细胞的上皮间质转化且促进细胞凋亡,miR-491负调控αB-crystallin是其关键分子机制之一。Objective To investigate the effect of miRNA-491(miR-491)on proliferation and apoptosis of human glioblastoma cells and its regulation mechanism.Methods Fresh glioblastoma multiforme tissue(GBM)samples and normal tissues(NT)adjacent to GBM were collected.Normal glial cell line(NHA)and glioblastoma cell lines U-87 MG and U-118 MG were cultured.U-118MG cells were divided into no treatment group(control group),miR-491-mimic group(mimic group),mimic negative control group(mimic-NC group),and mimic combined withαB-crystallin-expressed plasmid group(mimic+αB-crystallin group).The proliferation of cells was evaluated by CCK-8 method,and the expression levels of miR-491,αB-crystallin,Twist1,E-cadherin,N-cadherin,Bax and Bcl-2 were detected by RT-qPCR and Western blot.The luciferase reporter assay was used to validate the binding of miR-491 and the 3′UTR region ofαB-crystallin gene.Results The expression of miR-491 was significantly decreased in GBM tissues compared with NT(P<0.05),and it was decreased in U-87 MG and U-118 MG cells compared with NHA cells(P<0.05).Compared with control group,the expression levels ofαB-crystallin,N-cadherin and Twist1 were downregulated in mimic group,the apoptosis rate was up-regulated(P<0.05),and the level of E-cadherin was increased(P<0.05).Luciferase report confirmed the binding of miR-491 with the 3′UTR region ofαB-crystallin gene.In mimic+αB-crystallin group,the effects of mimic on the expression of apoptotic and epithelial-mesenchymal transition markers were reversed(P<0.05).Conclusion The miR-491 negatively regulatesαB-crystallin-mediated epithelial-mesenchymal transition and inflammatory response in glioblastoma cells.

关 键 词：胶质母细胞瘤 细胞上皮间质转化 细胞凋亡 miRNA-491 αB-crystallin 

分 类 号：R739.4[医药卫生—肿瘤]