【摘要】 目的:探讨肺腺癌相关转录本1(LUADT1)对胶质瘤细胞的增殖、迁移和侵袭的影响及分子机制。方法:将si-NC、si-LUADT1、miR-NC、miR-138-5p、pcDNA、pcDNA-LUADT1分别转染至U251细胞中,记为si-NC组、si-LUADT1组、miR-NC组、 miR-138-5p组、pcDNA组、pcDNA-LUADT1组;将si-LUADT1分别与anti-miR-NC、anti-miR-138-5p共转染至U251细胞中,记为si-LUADT1+anti-miR-NC组、si-LUADT1+anti-miR-138-5p组。实时荧光定量PCR(RT-qPCR)检测LUADT1和miR-138-5p表达水平;细胞计数试剂盒8(CCK-8)检测细胞增殖活性;Transwell检测细胞迁移和侵袭;蛋白质印迹(Western blot)法检测细胞周期蛋白D1(CyclinD1)、细胞周期蛋白依赖性激酶抑制剂1A(p21)、基质金属蛋白酶2(MMP-2)、基质金属蛋白酶9(MMP-9)的表达;双荧光素酶报告实验检测lncRNA LUADT1和miR-138-5p的靶向关系。结果:胶质瘤组织中lncRNA LUADT1高表达, miR-138-5p低表达。抑制lncRNALUADT1表达或过表达miR-138-5p,细胞活性降低,细胞迁移、侵袭数降低,CyclinD 1、MMP-2、MMP-9蛋白表达水平降低,p21蛋白表达水平升高。lncRNA LUADT1靶向调控miR-138-5p,干扰miR-138-5p表达逆转了抑制lncRNA LUADT1表达对胶质瘤U251细胞增殖、迁移、侵袭的抑制作用。结论:抑制lncRNA LUADT1表达通过靶向miR-138-5p抑制胶质瘤细胞的增殖、迁移和侵袭。更多还原

【Abstract】 Objective:To investigate the effects and molecular mechanisms of lung adenocarcinoma associated transcript 1(LUADT1) on the proliferation,migration and invasion of glioma cells.Methods:si-NC,si-LUADT1,miR-NC,miR-138-5 p,pcDNA and pcDNA-LUADT1 were transfected into U251 cells,and record as si-NC group,si-LUADT1 group,miR-NC group,miR-138-5 p group,pcDNA group and pcDNA-LUADT1 group.si-LUADT1 co-transfect with anti-miR-NC and anti-miR-138-5 p into U251 cells and record as si-LUADT1+anti-miR-NC group,si-LUADT1+anti-miR-138-5 p group.Real-time fluorescence quantitative PCR(RT-qPCR) detected the expressions of LUADT1 and miR-138-5 p.Cell counting kit 8(CCK-8) detected cell proliferation activity.Transwell detected cell migration and invasion.Western blot detected CyclinD1,cyclin-dependent kinase inhibitor 1 A(p21),matrix metalloproteinase 2(MMP-2),matrix metalloproteinase 9(MMP-9) protein expression.Dual luciferase reporting assay detected targeting relationship between LUADT1 and miR-138-5 p.Results:LUADT1 was highly expressed in glioma tissues and miR-138-5 p was low expressed.After inhibition of LUADT1 expression or overexpression of miR-138-5 p,cell viability was reduced,cell migration and invasion was reduced,CyclinD1,MMP-2,MMP-9 expressions were decreased,and p21 expression was increased.LUADT1 targeted miR-138-5 p,and interfering with miR-138-5 p expression reversed the inhibition of lncRNA LUADT1 expression on glioma U251 cell proliferation,migration,and invasion.Conclusion:Inhibiting the expression of LUADT1 can inhibit the proliferation,migration and invasion of glioma cells by targeting miR-138-5 p.更多还原