Wang Jianming, Ying Yulin, Bo Shumei, Li Guangjun, Yuan Fei
Department of Urology, Linyi People's Hospital, Linyi, Shandong 276003, P.R. China.
Department of Urology, Linshu People's Hospital, Linyi, Shandong 276700, P.R. China.
Mol Med Rep. 2016 Aug;14(2):1829-34. doi: 10.3892/mmr.2016.5403. Epub 2016 Jun 15.
The present study assessed the expression profiles of numerous microRNAs (miRs; miR-141, miR-187, miR-206, miR-218, miR-335 and miR-204) in 25 pairs of renal cell carcinoma (RCC) tissue samples and adjacent non-cancerous tissue. Reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) analysis revealed that miR‑218 was significantly downregulated in RCC tissue samples. Next, web‑based algorithms were used to identify B‑cell lymphoma (BCL)9 as a possible target of miR‑218, which was confirmed by a subsequent luciferase assay. Furthermore, the mRNA expression of BCL9 in the tumor and adjacent normal tissue samples was assessed, and BCL9 was markedly upregulated in the tumor tissue samples compared with the adjacent non‑cancerous controls. In addition, miR‑218 mimics or its inhibitors were transfected into renal cell carcinoma cells and harvested 48 h later. RT‑qPCR and western blotting revealed that both the mRNA and protein expression levels of BCL9 were significantly downregulated by the miR‑218 mimics. However, inhibition of miR‑218 upregulated the expression levels of BCL9. A 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) proliferation assay revealed that cell proliferation was suppressed in miR‑218 mimic‑transfected RCC cells compared with control cells; however, cell proliferation was significantly promoted in the RCC cells transfected with miR‑218 inhibitors compared with the controls. Taken together, it was demonstrated that miR‑218 modulated a novel molecular target and the present study provided novel insights into potential mechanisms of RCC oncogenesis.
本研究评估了25对肾细胞癌（RCC）组织样本及相邻非癌组织中多种微小RNA（miR；miR-141、miR-187、miR-206、miR-218、miR-335和miR-204）的表达谱。逆转录定量聚合酶链反应（RT-qPCR）分析显示，miR-218在RCC组织样本中显著下调。接下来，利用基于网络的算法将B细胞淋巴瘤（BCL）9鉴定为miR-218的一个可能靶点，随后的荧光素酶测定证实了这一点。此外，评估了肿瘤和相邻正常组织样本中BCL9的mRNA表达，与相邻非癌对照相比，肿瘤组织样本中BCL9明显上调。另外，将miR-218模拟物或其抑制剂转染到肾癌细胞中，48小时后收获细胞。RT-qPCR和蛋白质印迹分析显示，miR-218模拟物显著下调了BCL9的mRNA和蛋白质表达水平。然而，抑制miR-218上调了BCL9的表达水平。3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）增殖试验显示，与对照细胞相比，转染miR-218模拟物的RCC细胞中细胞增殖受到抑制；然而，与对照相比，转染miR-218抑制剂的RCC细胞中细胞增殖显著促进。综上所述，证明了miR-218调节了一个新的分子靶点，本研究为RCC肿瘤发生的潜在机制提供了新的见解。
