He Haowei, Wang Linhui, Zhou Wenquan, Zhang Zhengyu, Wang Longxin, Xu Song, Wang Dong, Dong Jie, Tang Chaopeng, Tang Hao, Yi Xiaoming, Ge Jingping
Department of Urology, Jinling Hospital, Nanjing University Medical School, Nanjing, Jiangsu, China.
Department of Urology, Changzheng Hospital, Second Military Medical University, Shanghai, Shanghai, China.
PLoS One. 2015 May 4;10(5):e0125672. doi: 10.1371/journal.pone.0125672. eCollection 2015.
This study aims to profile dysregulated microRNA (miRNA) expression in clear cell renal cell carcinoma (ccRCC) and to identify key regulatory miRNAs in ccRCC.
miRNA expression profiles in nine pairs of ccRCC tumor samples at three different stages and the adjacent, non-tumorous tissues were investigated using miRNA arrays. Eleven miRNAs were identified to be commonly dysregulated, including three up-regulated (miR-487a, miR-491-3p and miR-452) and eight down-regulated (miR-125b, miR-142-3p, miR-199a-5p, miR-22, miR-299-3p, miR-29a, miR-429, and miR-532-5p) in tumor tissues as compared with adjacent normal tissues. The 11 miRNAs and their predicted target genes were analyzed by Gene Ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis, and three key miRNAs (miR-199a-5p, miR-22 and miR-429) were identified by microRNA-gene network analysis. Dysregulation of the three key miRNAs were further validated in another cohort of 15 ccRCC samples, and the human kidney carcinoma cell line 786-O, as compared with five normal kidney samples. Further investigation showed that over-expression of miR-199a-5p significantly inhibited the invasion ability of 786-O cells. Luciferase reporter assays indicated that miR-199a-5p regulated expression of TGFBR1 and JunB by directly interacting with their 3' untranslated regions. Transfection of miR-199a-5p successfully suppressed expression of TGFBR1 and JunB in the human embryonic kidney 293T cells, further confirming the direct regulation of miR-199a-5p on these two genes.
This study identified 11 commonly dysregulated miRNAs in ccRCC, three of which (miR-199a-5p, miR-22 and miR-429) may represent key miRNAs involved in the pathogenesis of ccRCC. Further studies suggested that miR-199a-5p plays an important role in inhibition of cell invasion of ccRCC cells by suppressing expression of TGFBR1 and JunB.
本研究旨在分析透明细胞肾细胞癌（ccRCC）中失调的微小RNA（miRNA）表达谱，并鉴定ccRCC中的关键调控miRNA。
使用miRNA芯片研究了9对处于三个不同阶段的ccRCC肿瘤样本及其相邻的非肿瘤组织中的miRNA表达谱。鉴定出11种miRNA普遍失调，与相邻正常组织相比，肿瘤组织中有3种上调（miR-487a、miR-491-3p和miR-452），8种下调（miR-125b、miR-142-3p、miR-199a-5p、miR-22、miR-299-3p、miR-29a、miR-429和miR-532-5p）。通过基因本体论和京都基因与基因组百科全书（KEGG）通路富集分析对这11种miRNA及其预测的靶基因进行了分析，并通过miRNA-基因网络分析鉴定出3种关键miRNA（miR-199a-5p、miR-22和miR-429）。在另一组15个ccRCC样本以及人肾癌细胞系786-O中，与5个正常肾样本相比，进一步验证了这3种关键miRNA的失调情况。进一步研究表明，miR-199a-5p的过表达显著抑制了786-O细胞的侵袭能力。荧光素酶报告基因检测表明，miR-199a-5p通过直接与其3'非翻译区相互作用来调节TGFBR1和JunB的表达。在人胚肾293T细胞中，转染miR-199a-5p成功抑制了TGFBR1和JunB的表达，进一步证实了miR-199a-5p对这两个基因的直接调控作用。
本研究鉴定出ccRCC中11种普遍失调的miRNA，其中3种（miR-199a-5p、miR-22和miR-429）可能是参与ccRCC发病机制的关键miRNA。进一步研究表明，miR-199a-5p通过抑制TGFBR1和JunB的表达，在抑制ccRCC细胞的侵袭中发挥重要作用。
