miR-34b抑制非小细胞肺癌A549的侵袭和迁移  

miR-34b inhibits invasion and migration of non-small cell lung cancer A549 cells

作　　者：郭东明 张奇 何东升 江跃全[1] 王志强 GUO Dong-Ming;ZHANG Qi;HE Dong-Sheng;JIANG Yue-Qua;WANG Zhi-Qiang(Chongqing Key Laboratory of Translational Research for Cancer Metastasis and Individualized Treatment,Chongqing University Cancer Hospital&Chongqing Cancer Institute&Chongqing Cancer Hospital,Chongqing 400030,China)

机构地区：[1]重庆大学附属肿瘤医院/重庆市肿瘤研究所/重庆市肿瘤医院,肿瘤转移与个体化诊治转化研究重庆市重点实验室,重庆400030 [2]成都医学院第一附属医院心胸外科,成都610500

出　　处：《中国免疫学杂志》2018年第11期1679-1683,共5页Chinese Journal of Immunology

基　　金：重庆市卫生局资助项目(No.2013-2-122)。

摘　　要：目的:本研究旨在探索miR-34b对非小细胞肺癌A549侵袭和迁移的影响及HIF1α在其中所起的作用。方法:A549细胞转染miR-34b mimic和pc-HIF1α过表达载体以过表达miR-34b和缺氧诱导因子-1α(HIF1α)。A549细胞分为4组:A549(空对照)组,miR-34b mimic组,pc-HIF1α组和miR-34b mimic+pc-HIF1α组。实时定量PCR(q RT-PCR)检测miR-34b表达和HIF1α的m RNA水平。荧光素酶分析验证miR-34b和HIF1α的靶向关系。蛋白印迹检测HIF1α、基质金属蛋白酶2(MMP-2),Snail和血管内皮细胞生长因子(VEGF)的蛋白水平。Transwell分析细胞侵袭。划痕实验检测细胞迁移。结果:转染miR-34b mimic可提高A549细胞miR-34b的表达水平,降低HIF1α的m RNA水平(P <0. 001)。miR-34b可直接靶向HIF1α。miR-34b过表达可抑制pc-HIF1α诱导的HIF1α蛋白水平升高(P<0. 01)。与对照组相比,miR-34b过表达可显著降低细胞侵袭数目和愈合率(P<0. 01)。此外,miR-34b过表达可显著抑制HIF1α过表达导致的细胞侵袭和愈合率增加(P<0. 01)。miR-34b mimic组MMP-2、Snail和VEGF相对蛋白水平低于对照组(P<0. 01)。pc-HIF1α组MMP-2、Snail和VEGF相对蛋白水平高于对照组(P<0. 01)。与pc-HIF1α组相比,miR-34b mimic+pc-HIF1α组MMP-2、Snail和VEGF相对蛋白水平下降(P<0. 01)。结论:miR-34b-5p过表达通过靶向HIF1α抑制非小细胞肺癌A549的侵袭和迁移。Objective:To explore the effect of miR-34b on the invasion and migration of non-small cell lung cancer A549 cells and the role of HIF1α.Methods:Overexpression of miR-34b and HIF1αwas performed by transfection with miR-34b mimic and pc-HIF1αoverexpression vector in A549 cells.A549 cells were divided into four groups:A549(control)group,miR-34b mimic group,pc-HIF1αgroup and miR-34b mimic+pc-HIF1αgroup.The expression of miR-34b and mRNA level of HIF1αwas tested by quantitative real-time reverse transcription PCR(qRT-PCR).The targeted relationship of miR-34b and HIF1αwas verified by luciferase assay.The protein levels of HIF1α,matrix metalloprotein 2(MMP-2),Snail and vascular endothelial growth factor(VEGF)were measured by Western blot.Cell invasion was analysed by Transwell.Cell migration was detected by wound healing.Results:After transfection with miR-34b mimic in A549 cells,the expression of miR-34b was increased and the mRNA level of HIF1αwas decreased(P<0.001).HIF1αwas a direct target of miR-34b.Overexpression of miR-34b repressed the pc-HIF1α-induced elevated level of HIF1αprotein(P<0.01).Compared with control group,the number of invasive cells per field and wound closure rate in miR-34b mimic group were decreased(P<0.01).The number of invasive cells per field and wound closure rate in pc-HIF1αgroup were higher than control group(P<0.01).Compared with pc-HIF1αgroup,the number of invasive cells per field and wound closure rate in miR-34b mimic+pc-HIF1αgroup were reduced(P<0.01).The protein levels of MMP-2,Snail and VEGF in miR-34b mimic group were lower than control group(P<0.01).The protein levels of MMP-2,Snail and VEGF in pc-HIF1αgroup were higher than control group(P<0.01).Compared with pc-HIF1αgroup,the protein levels of MMP-2,Snail and VEGF in miR-34b mimic+pc-HIF1αgroup were decreased(P<0.01).Conclusion:miR-34b-5p overexpression suppresses the invasion and migration of non-small cell lung cancer A549 cells via targeting HIF1α.

关 键 词：肺癌 miR-34b-5p 侵袭 迁移 HIF1Α 

分 类 号：R734.2[医药卫生—肿瘤]