miR-200b通过靶向PROM1抑制胶质瘤细胞侵袭  

miR-200b suppresses glioma cell invasion by targeting PROM1

作　　者：彭彪[1] 胡辅 秦明筠[1] 罗冬冬[1] 张训[1] 赵海林[1] 

机构地区：[1]广州医科大学附属肿瘤医院神经外科,510095

出　　处：《中华肿瘤杂志》2015年第1期25-28,共4页Chinese Journal of Oncology

摘　　要：目的探讨miR-200b靶向调控PROM1的表达对胶质瘤细胞侵袭能力的影响以及miR-200b抑瘤的分子机制。方法构建PROM13’端非翻译区（3’UTR）荧光素酶报告载体，荧光素酶报告检测miR-200b对PROMI3’UTR-荧光素酶活性的影响。将miR-200b模拟物转染胶质瘤U87细胞，采用实时荧光定量PCR和Westernblot检测PROMlmRNA和蛋白的表达水平。将PROM1 siRNA转染U87细胞，Transwell侵袭实验检测PROM1下调对U87细胞侵袭能力的影响。结果双荧光素酶报告检测显示，miR．200b能特异性地与PROM13’UTR结合，抑制其荧光素酶活性，其荧光素酶活性强度下降了57．0％，差异有统计学意义（P〈0．01）。过表达miR-200b的U87细胞中PROM1蛋白和mRNA表达水平均降低，转染miR-200b组和对照组中PROMImRNA的表达水平分别为0．64±0．05和0．95±0．09，差异有统计学意义（P〈0．05）。siRNA干扰PROM1表达能抑制U87细胞的生长和侵袭能力。转染PROMIsiRNA组和对照组中穿膜细胞数分别为（85±9）个和（155±16）个，差异有统计学意义（P〈0．05）。结论miR．200b可通过靶向调控PROM1的表达而抑制胶质瘤细胞的侵袭力。Objective To explore whether miR-200b suppresses tumor cell invasion by targeting PROMI, thus to reveal the molecular mechanism that miR-200b functions as a tumor suppressor in glioma. Methods PROM13＇ UTR-luciferase vector was constructed and dual-luciferase reporter gene assay was employed to examine the effect of miR-200b on luciferase activity. Human glioblastoma U87 cells were transfected with miR-200b mimics, and next qRT-PCR and Western blotting were performed to detect the expressions of PROM1 mRNA and protein. The effect of PROMI down-regulation on invasion was observed after PROMI siRNA were transfected into U87 cells. Results The miR-200b bound to the 3＇-untranslated region （UTR） of PROMI and inhibited the luciferase activity. Its luciferase activity was down-regulated by 57.0% （P 〈 0.01 ）. PROM I protein and mRNA expressions were significantly down-regulated when miR- 200b was overexpressed in the U87 cells （ P 〈 0. 05 ）. siRNA-mediated down-regulation of PROM1 suppressed the potential of cell invasion. The invasion ability of SKOV3 cells after transfection with siRNA- PROM1 was significantly lower than that in the negative control cells （P 〈 0.05 ）. Conclusion miR-200b may suppress cell invasion by targeting PROM1 in glioma.

关 键 词：神经胶质瘤 肿瘤侵润 miR-200b PROM1 

分 类 号：R739.41[医药卫生—肿瘤]