Yao Kun, He Leye, Gan Yu, Zeng Qing, Dai Yingbo, Tan Jing
Department of Urology, The Third Xiangya Hospital of Central South University, 138 Tongzipo Road, Changsha, 410013, Hunan, China.
Diagn Pathol. 2015 Aug 20;10:146. doi: 10.1186/s13000-015-0372-3.
Increasing evidence has shown that microRNAs function as oncogenes or tumor suppressors in human malignancies, but the roles of miR-186 in human bladder cancer (BC) is still unclear.
First, quantitative real-time PCR (qRT-PCR) was performed to detect miR-186 expression in bladder cancer tissues and cell lines. Then, Bioinformatics analysis, combined with luciferase reporter assay demonstrated the target gene of miR-186. Finally, the roles of miR-186 in regulation of tumor proliferation and invasion were further investigated.
Here, our study showed miR-186 was down-regulated in bladder cancer tissues and cell lines. Luciferase reporter assay showed that miR-186 targets NSBP1 3'-untranslated region (UTR) directly and suppresses NSBP1 (HMGN5) expression in human bladder cancer cells. NSBP1 siRNA- and miR-186-mediated NSBP1 knock-down experiments revealed that miR-186 suppresses cell proliferation and invasion through suppression of NSBP1 expression. Expression analysis of a set of epithelial-mesenchymal transition (EMT) markers showed that NSBP1 involves miR-186 suppressed EMT which reducing the expression of mesenchymal markers (vimentin and N-cadherin) and inducing the expression of epithelial marker (E-cadherin).
Our data first time identified miR-186 as the upstream regulator of NSBP1 and also suggest miR-186-suppressed NSBP1 as a novel therapeutic approach for bladder cancer.
越来越多的证据表明,微小RNA在人类恶性肿瘤中发挥癌基因或肿瘤抑制因子的作用,但miR-186在人类膀胱癌(BC)中的作用仍不清楚。
首先,采用定量实时PCR(qRT-PCR)检测膀胱癌组织和细胞系中miR-186的表达。然后,通过生物信息学分析并结合荧光素酶报告基因检测确定miR-186的靶基因。最后,进一步研究miR-186在调节肿瘤增殖和侵袭中的作用。
在本研究中,我们发现miR-186在膀胱癌组织和细胞系中表达下调。荧光素酶报告基因检测表明,miR-186直接靶向NSBP1的3'-非翻译区(UTR),并抑制人膀胱癌细胞中NSBP1(HMGN5)的表达。NSBP1 siRNA和miR-186介导的NSBP1敲低实验表明,miR-186通过抑制NSBP1的表达来抑制细胞增殖和侵袭。一组上皮-间质转化(EMT)标志物的表达分析表明,NSBP1参与了miR-186抑制的EMT过程,即降低间质标志物(波形蛋白和N-钙黏蛋白)的表达并诱导上皮标志物(E-钙黏蛋白)的表达。
我们的数据首次确定miR-186为NSBP1的上游调节因子,并提示miR-186抑制的NSBP1可作为膀胱癌的一种新的治疗方法。