Role of pre-miR-532 (miR-532-5p and miR-532-3p) in regulation of gene expression and molecular pathogenesis in renal cell carcinoma.
Role of pre-miR-532 (miR-532-5p and miR-532-3p) in regulation of gene expression and molecular pathogenesis in renal cell carcinoma.
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DOI：
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发表时间：
2019
影响因子：
1.2
通讯作者：
Yasutaka Yamada;Takayuki Arai;Mayuko F Kato;S. Kojima;S. Sakamoto;A. Komiya;Y. Naya;T. Ichikawa;N. Seki
中科院分区：
文献类型：
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作者：
Yasutaka Yamada;Takayuki Arai;Mayuko F Kato;S. Kojima;S. Sakamoto;A. Komiya;Y. Naya;T. Ichikawa;N. Seki
对我们先前确定的肾细胞癌（RCC）的microRNA（miRNA）表达特征和癌症基因组图谱（TCGA）数据库的分析显示，前-miR-532-532- 5 p的两条链都是miR-532- 5 p。（引导链）和miR-532- 3 p（乘客链）-与肾细胞癌患者的不良预后密切相关（分别为P = 0.0411和P = 0.022）。在这项研究中，我们研究了这些miRNAs的功能意义，并确定了参与RCC发病机制的基因靶点。这些miRNAs的异位表达显著减弱了两种肾癌细胞系786-O和A498的恶性表型，包括增殖、迁移和侵袭。全基因组基因表达和计算机数据库分析的组合揭示了RCC细胞中分别由miR-532- 5 p和miR-532- 3 p调控的36和34个基因作为推定的靶癌基因。其中，水通道蛋白AQP 9的表达受miR-532- 5 p和miR-532- 3 p的直接调控，AQP 9高表达与肾癌患者预后不良显著相关（P = 2.03e-05）。多因素分析提示AQP 9表达是影响肾细胞癌患者预后的独立因素。肾细胞癌中AQP 9在基因和蛋白水平均存在异常表达。siRNA介导的si-AQP 9对肾癌细胞的恶性表型有抑制作用。AQP 9过表达的补救分析显示miR-532/AQP 9轴与RCC肿瘤发生密切相关。抗肿瘤miRNAs及其靶点的鉴定将有助于提高对RCC分子发病机制的理解。
Analyses of our previously determined microRNA (miRNA) expression signature of renal cell carcinoma (RCC) and The Cancer Genome Atlas (TCGA) database revealed that both strands of the pre-miR-532-duplex-miR-532-5p (the guide strand) and miR-532-3p (the passenger strand)- are closely associated with poor prognosis of RCC patients (P = 0.0411 and P = 0.022, respectively). In this study we investigated the functional significance of these miRNAs and identified gene targets involved in RCC pathogenesis. Ectopic expression of these miRNAs significantly attenuated the malignant phenotypes including proliferation, migration and invasion of two RCC cell lines, 786-O and A498. A combination of genome-wide gene expression and in silico database analyses revealed 36 and 34 genes as putative target oncogenes regulated by miR-532-5p and miR-532-3p, respectively, in RCC cells. Among these targets, expression of aquaporin9 (AQP9), a water channel protein, was directly regulated by both miR-532-5p and miR-532-3p, and high expression levels of AQP9 were significantly associated with poor prognosis of RCC patients (P = 2.03e-05). Multivariate analysis indicated that AQP9 expression is an independent prognostic factor for RCC patients. Aberrant AQP9 expression at both the gene and protein level was detected in RCC clinical specimens. siRNA-mediated knockdown of AQP9 by si-AQP9 inhibited the malignant phenotypes of RCC cells. Rescue assays of AQP9 overexpression showed that the miR-532/AQP9 axis was closely involved in RCC oncogenesis. The identification of antitumor miRNAs and their targets will contribute to an increased understanding of the molecular pathogenesis of RCC.