miR-3941抑制securin的表达对神经胶质瘤细胞的影响  

Effects of miR-3941 on glioma cells by inhibiting securin expression

作　　者：闫海成[1] 苏乌云[2] 李玉莲[3] 都兰 陈秀兰 刘淑娟 王薇[2] YAN Hai-cheng;SU Wu-yun;LI Yu-lian;DU Lan;CHEN Xiu-lan;LIU Shu-juan;WANG Wei(Department of Neurosurgery,Affiliated Hospital of Inner Mongolia Medical University,Hohhot,Inner Mongolia 010010,China;Department of Medical Oncology,Affiliated Hospital of Inner Mongolia Medical University,Hohhot,Inner Mongolia 010010,China;Department of Pathology,Affiliated Hospital of Inner Mongolia Medical University,Hohhot,Inner Mongolia 010010,China)

机构地区：[1]内蒙古医科大学附属医院神经外科,内蒙古自治区呼和浩特010010 [2]内蒙古医科大学附属医院肿瘤内科,内蒙古自治区呼和浩特010010 [3]内蒙古医科大学附属医院病理科,内蒙古自治区呼和浩特010010

出　　处：《热带医学杂志》2022年第11期1461-1466,1595,1456,共8页Journal of Tropical Medicine

基　　金：国家自然科学基金(81660469);内蒙古自然科学基金(2020LH08035);希思科-石药肿瘤研究基金(Y-sy2018-134)

摘　　要：目的探讨miR-3941对神经胶质瘤发生发展的影响以及可能的分子机制。方法为了明确miR-3941的功能,本研究将细胞分为三组,一组不做任何处理(U251细胞组),一组转染NC mimics作为对照组(NC mimics组),一组转染miR-3941 mimics(miR-3941 mimics组)。为了研究securin对miR-3941功能的影响,又将细胞分为三组,一组转染miR-3941 mimics(miR-3941 mimics组),一组共转染miR-3941 mimics和空质粒作为对照(miR-3941mimics+NC组),一组共转染miR-3941 mimics和securin过表达质粒(miR-3941 mimics+ov-securin组)。采用RTqPCR检测人正常胶质细胞和胶质瘤U251细胞中miR-3941的表达情况。CCK-8实验和Transwell实验检测过表达miR-3941、过表达miR-3941同时过表达securin对胶质瘤U251细胞增殖、迁移和侵袭影响。Western blot检测过表达miR-3941、过表达miR-3941同时过表达securin对胶质瘤U251细胞中securin蛋白表达的影响。双荧光素酶报告实验分析miR-3941对securin的靶向作用关系。结果与人正常胶质细胞相比,胶质瘤U251细胞中miR-3941相对表达水平显著下调,差异有统计学意义(t=57.42,P<0.05)。CCK-8实验表明在培养48和72 h时U251细胞组、NC mimics组以及miR-3941 mimics组之间细胞增殖差异有统计学意义(F=424.50、956.20,P均<0.05);miR-3941 mimics组的增殖明显低于NC mimics组(P<0.05)。Transwell实验表明U251细胞组、NC mimics组以及miR-3941 mimics组的细胞迁移和侵袭差异有统计学意义(F=6361.00、5608.00,P均<0.05);和NC mimics组相比,转染miR-3941mimics组迁移和侵袭细胞数显著减少(P<0.05)。双荧光素酶实验结果显示,与野生型securin 3’-UTR报告载体共转染后,U251组、miR-3941 mimics组和miR-3941 inhibitor组之间荧光素酶活性差异有统计学意义(F=649.10,P<0.05),而与突变后的securin 3’-UTR报告载体共转染后,以上三组荧光强度无明显变化,差异无统计学意义(F=0.55,P=0.605)。Western blot检测发现转染miR-3941 mimics的胶质瘤U251�Objective To investigate the effects of miR-3941 on the occurrence and development of glioma and its possiblemolecular mechanism.MethodsTo clarify the function of mi R-3941,the cells were divided into three groups in this study.One group without any treatment(U251 cell group),one group was transfected with NC mimics as control group(NCmimics),and one group with mi R-3941 mimics(mi R-3941 mimics group).In addition,to investigate the effect of securin on miR-3941 function,the cells were divided into three groups.One group was transfected with miR-3941 mimics(mi R-3941 mimics group),one group cotransfected with mi R-3941 mimics and empty plasmid as a control(mi R-3941mimics+NC group),and one group cotransfected with mi R-3941 mimics and securin overexpression plasmids(mi R-3941mimics+ov-securin group).The expression of mi R-3941 in human normal glial cells and glioma U251 cells was measured byRT-q PCR.The effects of mi R-3941 overexpression,both mi R-3941 and securin overexpression on the proliferation,migration,and invasion of glioma U251 cells were detected by CCK-8 assay and Transwell assay,respectively.The effectsof overexpression of mi R-3941,both mi R-3941 and securin overexpression on the expression of securin in glioma U251 cellswas measured by Western blot.The targeting effect of mi R-3941 on securin was evaluated by Dual luciferase reporter assay.ResultsCompared with human normal glial cells,the relative expression level of mi R-3941 in glioma U251 cells wassignificantly down regulated(t=57.42,P<0.05).CCK-8 assay showed that the proliferation in U251 cells,NC mimics,andmi R-3941 mimics groups was significantly different at 48 and 72 h(F=424.50,956.20,all P<0.05).The proliferation ofglioma U251 cells transfected with mi R-3941 mimics was significantly lower than that of the NC mimics group at 48 and 72 h(P<0.05).Transwell assay showed that migration and invasion were significantly different between the U251 cell group,NCmimics and mi R-3941 mimics groups(F=6361.00,5608.00,all P<0.05).Compared with the NC mi

关 键 词：胶质瘤 miR-3941 增殖 迁移 侵袭 

分 类 号：R739.41[医药卫生—肿瘤]