miR-503-5p靶向bFGF对人膀胱癌T24细胞侵袭和迁移的调节作用  

Modulation of miR-503-5p on invasion and migration of human bladder cancer T24 cells by targeting bFGF

作　　者：刘昌明[1] 黄香尘 杜斌[1] LIU Chang-Ming;HUANG Xiang-Chen;DU Bin(Department of Pathological,Ya′an Vocational and Technical College,Ya′an 625000,China)

机构地区：[1]雅安职业技术学院病理教研室,雅安625000 [2]雅安职业技术学院附属医院病理科,雅安625000

出　　处：《中国免疫学杂志》2019年第21期2618-2622,2627,共6页Chinese Journal of Immunology

基　　金：四川省医学青年科研计划项目(Q15008)

摘　　要：目的:本文主要研究miR-503-5p对膀胱癌细胞T24侵袭和迁移的调节作用。方法:首先,通过基因预测软件TargetScan和miRanda筛选出miR-503-5p的靶基因,并通过荧光素酶报告实验检测;然后,miR-503-5p mimic和pcDNA-bFGF单独或联合转染膀胱癌细胞T24,RT-PCR检测miR-503-5p和bFGF的表达,Transwell检测细胞侵袭情况,划痕实验检测细胞迁移能力,Western blot检测β-catenin、E-cadherin和N-cadherin的表达,接着,裸鼠皮下注射转染了miR-503-5p mimic的膀胱癌细胞T24构建移植瘤模型,30 d后检测肿瘤重量,RT-PCR检测miR-503-5p和bFGF的表达,免疫组化检测Ki67和β-catenin。结果:miR-503-5p与bFGF在3′UTR区存在结合位点,并且miR-503-5p直接靶向作用于bFGF。在体外,miR-503-5p抑制bFGF表达,miR-503-5p mimic组与Control组相比,侵袭细胞数目显著减少,迁移速率显著降低,E-cadherin表达显著上调,β-catenin和N-cadherin表达显著下调。在体内,miR-503-5p mimic组与Control组相比,肿瘤重量显著减轻,抑制bFGF的表达,Ki67和β-catenin阳性比率显著减少。结论:过表达miR-503-5p通过靶向抑制bFGF表达抑制膀胱癌细胞T24细胞的侵袭、迁移和上皮-间充质转化,从而抑制膀胱癌。Objective:To investigate the regulation of miR-503-5p on invasion and migration of bladder cancer cell line T24.Methods: First,the target genes of miR-503-5p were screened by the gene prediction software TargetScan and miRanda,and detected by luciferase reporter assay.Then,bladder cancer T24 cells were transfected alone or in combination by miR-503-5p mimic and pcDNA-bFGF.In vitro,the expression of miR-503-5p and bFGF was detected by RT-PCR,the cell invasion was detected by Transwell,the cell migration ability was detected by scratch test,and β-catenin,E-cadherin and N-cadherin were detected by Western blot.In vivo,transfected nude mice with subcutaneous injection of miR-503-5p mimic bladder cancer cell T24 construct model,tumor weight was measured after 30 days,the expression of miR-503-5p and bFGF was detected by RT-PCR,Ki67 and β-catenin were detected by immunohistochemistry.Results: miR-503-5p and bFGF had binding sites in the 3′UTR region,and miR-503-5p directly targets bFGF.In vitro,miR-503-5p inhibits bFGF expression,miR-503-5p mimic group compared with the Control group,the number of invasive cells was significantly decreased,the migration rate was significantly decreased,the expression of E-cadherin was significantly up-regulated,and the expression of β-catenin and N-cadherin was significantly down-regulated.In vivo,the miR-503-5p mimic group was compared with the Control group.Tumor weight was significantly reduced,miR-503-5p inhibition of bFGF expression,and a significant decrease in Ki67 and β-catenin positive rates.Conclusion: Overexpression of miR-503-5p inhibits bladder cancer T24 cell invasion,migration and epithelial-mesenchymal transition by targeting inhibition of bFGF,thereby inhibiting bladder cancer.

关 键 词：miR-503-5p BFGF 膀胱癌细胞T24 侵袭 迁移 

分 类 号：R734.2[医药卫生—肿瘤]