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微小RNA-320a下调通过靶向整合素β3介导膀胱癌侵袭。

MiR-320a down-regulation mediates bladder carcinoma invasion by targeting ITGB3.

作者信息

Shang Chao, Zhang Hui, Guo Yan, Hong Yang, Liu Yunhui, Xue Yixue

机构信息

Department of Neurobiology, China Medical University, Shenyang, 110001, People's Republic of China.

出版信息

Mol Biol Rep. 2014;41(4):2521-7. doi: 10.1007/s11033-014-3110-0. Epub 2014 Jan 18.

Abstract

To investigate whether the miR-320a could regulate bladder cancer cells invasion by down-regulation of ITGB3. Real-time quantitative PCR was applied to evaluate the expression level of miRNA-320a in bladder transitional cell carcinomas (TCC) and normal bladder transitional cell (NBTC) samples. The invasion ability of miR-320a in TCC T24 cells was analyzed by Transwell assay after pre-miR-320a or anti-miR-125b transfection. For the invasion mechanism analysis of miR-320a on T24 cells, TargetScan, PicTar and miRBase were used to predict the possible target gene of miR-320a. Luciferase activities assay and western blot were used to reveal the predicted target gene of miR-320a were direct and specific. RNA interference technology was used to confirm the invasion inhibition of miR-320a was directly induced by ITGB3. Our study showed that miR-320a was down-regulated in human TCC specimens compared to that in NBTC specimens. Over-expression of miR-320a in T24 cells inhibited TCC invasion and this inhibitory effect on T24 cells could be restore by miR-320a knocked down. Mechanism analysis revealed that ITGB3 was a direct and specific target of miR-320a. The advanced effect of anti-miR-320a on TCC cell invasion was mediated by expression silence of ITGB3. In summary, aberrantly expressed miR-320a contribute to T24 cells invasion partly through directly down-regulating ITGB3 protein expression in TCC and this miRNA signature offers a novel potential therapeutic strategy for TCC.

摘要

为研究miR-320a是否可通过下调整合素β3(ITGB3)来调节膀胱癌细胞的侵袭。应用实时定量聚合酶链反应(PCR)评估miRNA-320a在膀胱移行细胞癌(TCC)和正常膀胱移行细胞(NBTC)样本中的表达水平。在转染前体miR-320a或抗miR-125b后,通过Transwell实验分析miR-320a对TCC T24细胞侵袭能力的影响。为分析miR-320a对T24细胞的侵袭机制,使用TargetScan、PicTar和miRBase预测miR-320a可能的靶基因。通过荧光素酶活性测定和蛋白质免疫印迹法(western blot)揭示miR-320a预测的靶基因是直接且特异的。采用RNA干扰技术证实miR-320a对侵袭的抑制作用是由ITGB3直接诱导的。我们的研究表明,与NBTC标本相比,miR-320a在人类TCC标本中表达下调。T24细胞中miR-320a的过表达抑制了TCC的侵袭,而这种对T24细胞的抑制作用可通过敲低miR-320a来恢复。机制分析显示,ITGB3是miR-320a直接且特异的靶标。抗miR-320a对TCC细胞侵袭的促进作用是由ITGB3的表达沉默介导的。总之,异常表达的miR-320a部分通过直接下调TCC中ITGB3蛋白的表达促进T24细胞的侵袭,并且这种微小RNA特征为TCC提供了一种新的潜在治疗策略。

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