miR-513a-3p靶向鼠双微体基因2对胃癌细胞增殖迁移侵袭的影响  

Effect of miR-513a-3p targeting MDM2 on proliferation,migration and invasion of gastric cancer cells

作　　者：马颖光[1] 张自森[1] 于泳[1] 许晓芳[1] 袁丽[1] Ma Yingguang;Zhang Zisen;Yu Yong;Xu Xiaofang;Yuan Li(Department of Gastroenterology,the Fifth Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China)

机构地区：[1]郑州大学第五附属医院消化内科,450052

出　　处：《中华肿瘤杂志》2020年第1期30-36,共7页Chinese Journal of Oncology

基　　金：河南省医学科技攻关计划(201701015)。

摘　　要：目的探讨miR-513a-3p靶向鼠双微体基因2(MDM2)对胃癌细胞的增殖、迁移和侵袭的影响及其作用机制。方法采用脂质体法将miR-NC、miR-513a-3p、anti-miR-NC、anti-miR-513a-3p、si-NC、si-MDM2、miR-513a-3p+pcDNA3.1和miR-513a-3p+pcDNA3.1-MDM2转染至BGC-823细胞中,采用实时荧光定量聚合酶链反应(qRT-PCR)检测miR-513a-3p的表达水平,采用Western blot检测cyclin D1、MMP-2、p21、E-cadherin和MDM2蛋白的表达水平,四甲基偶氮唑蓝法检测各组胃癌细胞BGC-823的活性,Transwell法检测各组胃癌细胞BGC-823的迁移和侵袭能力,双荧光素酶报告基因检测实验检测miR-513a-3p与MDM2的靶向关系。结果胃癌细胞BGC-823、MGC-803中miR-513a-3p的表达水平分别为0.21±0.02和0.34±0.03,与胃上皮细胞GES-1(0.76±0.08)比较,差异均有统计学意义(均P<0.05)。培养24、48和72 h后,miR-NC组细胞的吸光度(A)值分别为0.57±0.05、1.03±0.10和1.43±0.14,miR-513a-3p组细胞的A值分别为0.36±0.03、0.48±0.05和0.63±0.06,差异均有统计学意义(均P<0.05);miR-NC组细胞的迁移和侵袭数分别为(130±11.80)个和(117±10.60)个,miR-513a-3p组细胞分别为(58±5.64)个和(50±5.13)个,差异均有统计学意义(均P<0.05)。培养24、48和72 h后,si-NC组细胞的A值分别为0.53±0.05、0.95±0.10和1.36±0.14,si-MDM2组细胞的A值分别为0.39±0.04、0.57±0.06和0.80±0.08;si-NC组细胞的迁移和侵袭数分别为(141±12.02)个和(109±10.60)个,si-MDM2组的迁移和侵袭数分别为(66±6.67)个和(61±6.18)个,差异均有统计学意义(均P<0.05)。培养24、48和72 h后,miR-513a-3p+pcDNA3.1组细胞的A值分别为0.34±0.03、0.46±0.05和0.61±0.06,miR-513a-3p+pcDNA3.1-MDM2组细胞的A值分别为0.48±0.05、0.82±0.08和1.17±0.12,差异均有统计学意义(均P<0.05);miR-513a-3p+pcDNA3.1组细胞的迁移和侵袭数分别为(56±5.71)个和(51±5.16)个,miR-513a-3p+pcDNA3.1-MDM2组分别为(113±10.28)个和(104±10.02)个,差异均有统计学意义(均P<0.05)。结�Objective To investigate the effects of miR-513a-3p on proliferation,migration and invasion of gastric cancer cells and its mechanism.Methods The miR-NC(miR-negative control mimics),miR-513a-3p(miR-513a-3p mimics),anti-miR-NC,anti-miR-513a-3p,si-NC,si-MDM2(murine double minute 2),miR-513a-3p+pcDNA3.1(co-transfected with miR-513a-3p and pcDNA3.1),miR-513a-3p+pcDNA3.1-MDM2(co-transfected with miR-513a-3p and pcDNA3.1-MDM2)were transfected into BGC-823 cells,respectively.The expression of miR-513a-3p was detected by real-time quantitative reverse transcription polymerase chain reaction(qRT-PCR),and the protein expressions of cyclin D1,MMP-2,p21,E-cadherin,MDM2 were detected by western blot.The viability of BGC-823 cells of each group was detected by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide(MTT)assay.The migration and invasion of each group were detected by Transwell,the targeting relationship between miR-513a-3p and MDM2 was detected by double luciferase reporter gene assay.Results The expression of miR-513a-3p in gastric epithelial cells GES-1 was 0.76±0.08,significantly higher than 0.21±0.02 in gastric cancer cells BGC-823 and 0.34±0.03 in MGC-803,respectively(P<0.05).The cell viabilities of the miR-NC group at 24 h,48 h and 72 h were 0.57±0.05,1.03±0.10,1.43±0.14,respectively,while those of the miR-513a-3p group were 0.36±0.03,0.48±0.05,and 0.63±0.06,respectively.The migration and invasion numbers of miR-NC group were 130±11.80 and 117±10.60,respectively,those of miR-513a-3p group were 58±5.64 and 50±5.13,respectively,and the differences were statistically significant(P<0.05).The cell viabilities of the si-NC group at 24 h,48 h and 72 h were 0.53±0.05,0.95±0.10,1.36±0.14,respectively.Those of the si-MDM2 group were 0.39±0.04,0.57±0.06,and 0.80±0.08,respectively.The cell migration and invasion of the si-NC group were 141±12.02 and 109±10.60,respectively,while those of the MDM2 group were 66±6.67 and 61±6.18,respectively,and the differences were statistically signific

关 键 词：胃肿瘤 miR-513a-3p 鼠双微体基因2 增殖 迁移 侵袭 

分 类 号：R735[医药卫生—肿瘤]