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长链非编码RNA PCAT1通过海绵化miR-656和miR-539上调YAP,从而驱动肾透明细胞癌。

Long non-coding RNA PCAT1 drives clear cell renal cell carcinoma by upregulating YAP via sponging miR-656 and miR-539.

作者信息

Wang Rui, Zheng Bin, Liu Hongyan, Wan Xiuxian

机构信息

Division of Nephrology, Renmin Hospital of Wuhan University, Wuhan, China.

Department of Nephrology, The Affiliated Lianyungang Oriental Hospital of Xuzhou Medical University, Lianyungang, Jiangsu, China.

出版信息

Cell Cycle. 2020 May;19(10):1122-1131. doi: 10.1080/15384101.2020.1748949. Epub 2020 Apr 14.

Abstract

Clear cell renal cell carcinoma (ccRCC) is the most common RCC subtype with high metastasis, poor prognosis and conventional chemotherapy resistance. Prostate cancer associated transcript 1 (PCAT1) is an important lncRNA that was reported to be involved in cell proliferation, migration and invasion of several types of cancer cells. However, its role in ccRCC is still undetermined. This study found that PCAT1 levels were elevated in ccRCC tumors as well as several ccRCC cells, and knockdown of PCAT1 with siRNA (si-PCAT1) alleviated cell proliferation, migration and invasion of Caki-2 and ACHN cells. With bioinformatics analysis, dual-luciferase reported assay, RNA pull-down assay and Spearman's correlation analysis, we demonstrated that PCAT1 acted as a sponge for miR-656 and miR-539. Moreover, we found dual competitive interaction of miR-656/539 with PCAT1 and yes-associated protein (YAP), resulting in the identification of PCAT1-miR-656/539-YAP axis in Caki-2 and ACHN cells. With CCK-8 assay and transwell assay, miR-656/539 inhibitor or YAP overexpression could alleviate the effects of si-PCAT1 on the proliferation, migration and invasion of Caki-2 and ACHN cells. Our data indicated that PCAT1 promotes proliferation, migration and invasion of ccRCC cells by upregulating YAP via sponging miR-656 and miR-539. Taken together, this study provided a novel therapeutic target for ccRCC treatment.

摘要

透明细胞肾细胞癌(ccRCC)是最常见的肾细胞癌亚型,具有高转移率、预后差和对传统化疗耐药的特点。前列腺癌相关转录本1(PCAT1)是一种重要的长链非编码RNA(lncRNA),据报道它参与多种癌细胞的细胞增殖、迁移和侵袭过程。然而,其在ccRCC中的作用仍未明确。本研究发现,PCAT1在ccRCC肿瘤以及几种ccRCC细胞中表达水平升高,用小干扰RNA(siRNA)敲低PCAT1(si-PCAT1)可减轻Caki-2和ACHN细胞的增殖、迁移和侵袭能力。通过生物信息学分析、双荧光素酶报告基因检测、RNA下拉实验和Spearman相关性分析,我们证明PCAT1作为miR-656和miR-539的海绵发挥作用。此外,我们发现miR-656/539与PCAT1和Yes相关蛋白(YAP)之间存在双重竞争性相互作用,从而在Caki-2和ACHN细胞中确定了PCAT1-miR-656/539-YAP轴。通过CCK-8实验和Transwell实验,miR-656/539抑制剂或YAP过表达可减轻si-PCAT1对Caki-2和ACHN细胞增殖、迁移和侵袭的影响。我们的数据表明,PCAT1通过海绵吸附miR-656和miR-539上调YAP,从而促进ccRCC细胞的增殖、迁移和侵袭。综上所述,本研究为ccRCC治疗提供了一个新的治疗靶点。

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