Peng Biao, Hu Su, Qin Mingjun, Luo Dongdong, Zhang Xun, Zhao Hailin
Department of Neurosurgery, the Affilated Cancer Hospital, Guangzhou Medical University. Guangzhou 510095, China. Email:
Department of Neurosurgery, the Affilated Cancer Hospital, Guangzhou Medical University. Guangzhou 510095, China.
Zhonghua Zhong Liu Za Zhi. 2015 Jan;37(1):25-8.
To explore whether miR-200b suppresses tumor cell invasion by targeting PROM1, thus to reveal the molecular mechanism that miR-200b functions as a tumor suppressor in glioma.
PROM1 3'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 PROM1 down-regulation on invasion was observed after PROM1 siRNA were transfected into U87 cells.
The miR-200b bound to the 3'-untranslated region (UTR) of PROM1 and inhibited the luciferase activity. Its luciferase activity was down-regulated by 57.0% (P < 0.01). PROM1 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).
miR-200b may suppress cell invasion by targeting PROM1 in glioma.
探讨miR-200b是否通过靶向PROM1抑制肿瘤细胞侵袭，从而揭示miR-200b在胶质瘤中作为肿瘤抑制因子发挥作用的分子机制。
构建PROM1 3'UTR荧光素酶载体，采用双荧光素酶报告基因检测法检测miR-200b对荧光素酶活性的影响。将miR-200b模拟物转染到人胶质母细胞瘤U87细胞中，然后进行qRT-PCR和蛋白质印迹法检测PROM1 mRNA和蛋白的表达。将PROM1 siRNA转染到U87细胞中后，观察PROM1下调对侵袭的影响。
miR-200b与PROM1的3'-非翻译区（UTR）结合并抑制荧光素酶活性。其荧光素酶活性下调了57.0%（P<0.01）。当miR-200b在U87细胞中过表达时，PROM1蛋白和mRNA表达明显下调（P<0.05）。siRNA介导的PROM1下调抑制了细胞侵袭潜能。用siRNA-PROM1转染后SKOV3细胞的侵袭能力明显低于阴性对照细胞（P<0.05）。
miR-200b可能通过靶向胶质瘤中的PROM1抑制细胞侵袭。
