miRNA-149靶向IGFBP5抑制膀胱癌细胞侵袭的分子机制研究  

Molecular mechanism of miRNA-149 targeting IGFBP5 to inhibit invasion of bladder cancer cells

作　　者：饶大庞[1] 虞海峰[1] 王帅彬[1] 孙来芳[1] RAO Dapang;YU Haifeng;WANG Shuaibin;SUN Laifang(Department of Urology,Yuying Children's Hospital,Second Affiliated Hospital of Wenzhou Medical University,Wenzhou 325000,China)

机构地区：[1]温州医科大学附属第二医院育英儿童医院泌尿外科,浙江温州325000

出　　处：《中国现代医生》2018年第35期1-4,14,169,共6页China Modern Doctor

基　　金：浙江省公益技术研究社会发展项目(2015C33216)

摘　　要：目的探讨miRNA-149靶向IGFBP5抑制膀胱癌细胞侵袭的分子机制。方法采用双萤光报告基因系统分别检测miRNA-149对目的基因IGFBP5和PDGFRA的3'UTR的影响。在T24膀胱癌细胞中瞬时转染miRNA-149模拟物(mimic)和阴性对照miRNA(mimic NC),并且在该细胞中分别转染IGFBP5过表达质粒(H10495)、阴性对照(H155)和PDGFRA过表达质粒(H10496),利用Transwell法检测细胞的迁移和侵袭能力。结果 miRNA-149显著抑制IGFBP5的3’UTR活性,差异具有统计学意义(P<0.0001)。Transwell实验显示T24-H155-miRNA149-mimics组较T24-H155-mimic-NC对照组(control)侵袭能力降低,差异具有统计学意义(P<0.0001),而在T24-H10495-miRNA-149-mimics组较T24-H155-miRNA-149-mimics组侵袭能力明显增强(P<0.01)。另一方面,T24-H10496-miRNA-149-mimics组较T24-H155-miRNA-149-mimics组侵袭能力无明显变化(P>0.05)。结论该研究表明miRNA-149通过抑制其下游基因IGFBP5从而抑制膀胱癌侵袭,为临床膀胱癌转移的分子治疗研究提供了新的靶点和理论依据。Objective To investigate the molecular mechanism of miRNA-149 targeting IGFBP5 to inhibit the invasion of bladder cancer cells.Methods The effect of miRNA-149 on the 3'UTR of the target genes IGFBP5 and PDGFRA was examined using a dual fluorescent reporter system.miRNA-149 mimics and negative control miRNAs(mimic NC)were transiently transfected into T24 bladder cancer cells,and IGFBP5 overexpression plasmid(H10495),negative control(H155)and PDGFRA overexpression plasmid(H10496)were transfected into the cells,respectively.The migration and invasion ability of the cells were detected by Transwell method.Results miRNA-149 significantly inhibited the 3'UTR activity of IGFBP5 with a statistically significant difference(P<0.0001).Transwell experiment showed that the T24-H155-miRNA149-mimics group had a lower invasive ability than the T24-H155-mimic-NC control group,and the difference was significant(P<0.0001).While the invasive ability of the T24-H10495-miRNA-149-mimics group was significantly enhanced compared with that of the T24-H155-miRNA-149-mimics group(P<0.0001).On the other hand,there was no significant change in the invasive ability of the T24-H10496-miRNA-149-mimics group compared with that of the T24-H155-miRNA-149-mimics group(P>0.05).Conclusion This study demonstrates that miRNA-149 inhibits the invasion of bladder cancer through inhibiting its downstream gene IGFBP5,which provides a new target and theoretical basis for the molecular treatment of clinical bladder cancer metastasis.

关 键 词：miRNA-149 膀胱癌 IGFBP5 PDGFRA 侵袭 

分 类 号：R737.9[医药卫生—肿瘤]