MiR-296-3p regulates cell growth and multi-drug resistance of human glioblastoma by targeting ether-a-go-go (EAG1)
MiR-296-3p regulates cell growth and multi-drug resistance of human glioblastoma by targeting ether-a-go-go (EAG1)
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DOI：
10.1016/j.ejca.2012.08.020
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发表时间：
2013-02-01
影响因子：
8.4
通讯作者：
Guo, Linlang
中科院分区：
文献类型：
--
作者：
Bai, Yifeng;Liao, Hongzhan;Guo, Linlang
microRNA（miRNAs）是一类短的非编码RNA分子，在转录后调节基因表达，在发育、分化、凋亡和增殖等多种生物学过程中发挥重要作用。为了探讨miRNAs在胶质母细胞瘤多药耐药（MDR）发生中的作用，我们首先采用Western blotting、实时荧光定量PCR（RT-PCR）和免疫组织化学方法检测了miR-296- 3 p和ether-a-go-go（EAG 1或KCNH 1）在U251细胞、U251/imatinib mesylate（U251 AR细胞）和临床标本中的表达。结果显示，与亲本U251细胞相比，U251 AR细胞中miR-296- 3 p表达下调，同时EAG 1蛋白表达上调。体外药物敏感性实验表明，过表达miR-296- 3 p可使胶质母细胞瘤（GBM）细胞对抗癌药物敏感，而使用反义寡核苷酸下调miR-296- 3 p表达可使GBM细胞产生MDR。miR-296- 3 p的异位表达降低了U251 AR细胞中EAG 1的表达并抑制了细胞增殖、耐药性和基于EAG 1 30非翻译区的报告基因构建体的荧光素酶活性，而EAG 1过表达则挽救了miR-296- 3 p在U251 AR细胞中的抑制作用。我们还发现EAG 1在临床标本中广泛过表达，并与miR-296- 3 p呈负相关。综上所述，我们的研究结果表明，miR-296- 3 p可能至少部分通过靶向EAG 1在胶质母细胞瘤中发挥MDR作用。(C)2012爱思唯尔有限公司保留所有权利。
MicroRNAs (miRNAs)-short non-coding RNA molecules-post-transcriptionally regulate gene expressions and play crucial roles in diverse biological processes such as development, differentiation, apoptosis and proliferation. In order to investigate the possible role of miRNAs in the development of multi-drug resistance (MDR) in human glioblastoma, we first detected (by Western blotting, real-time polymerase chain reaction [RT-PCR] and immunohistochemistry) the expression of miR-296-3p and ether-a-go-go (EAG1 or KCNH1) in U251 cells, U251/imatinib mesylate (U251AR cells) and clinical specimens. The results showed that miR-296-3p was down-regulated in U251AR cells, concurrent with the up-regulation of EAG1 protein, compared with the parental U251 cell line. In vitro drug sensitivity assay demonstrated that over-expression of miR-296-3p sensitised glioblastoma (GBM) cells to anticancer drugs, whereas down-expression using antisense oligonucleotides conferred MDR. Ectopic expression of miR-296-3p reduced EAG1 expression and suppressed cell proliferation drug resistance, and the luciferase activity of an EAG1 30-untranslated region-based reporter construct in U251AR cells, whereas EAG1 over-expression rescued the suppressive effect of miR-296-3p in U251AR cells. We also found that EAG1 was widely over-expressed and inversely correlated with miR-296-3p in clinical specimens. Taken together, our findings suggest that miR-296-3p may play a role of MDR in glioblastoma at least in part by targeting EAG1. (C) 2012 Elsevier Ltd. All rights reserved.