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长链非编码RNA通过EZH2促进增殖并预测透明细胞肾细胞癌的不良预后。

Long noncoding RNA promotes proliferation through EZH2 and predicts poor prognosis in clear cell renal cell carcinoma.

作者信息

Duan Junyao, Ma Xin, Shi Jing, Xuan Yundong, Wang Hanfeng, Li Pin, Zhang Yu, Fan Yang, Gong Huijie, Ma Xuetao, Pang Yuewen, Wang Ling, Yan Yongji, Zhang Xu

机构信息

Department of Urology, Dongzhimen Hospital Affiliated to Beijing University of Chinese Medicine, Beijing 100700, People's Republic of China.

Department of Urology, State Key Laboratory of Kidney Diseases, PLA Medical School, Chinese People's Liberation Army General Hospital, Beijing 100853, People's Republic of China.

出版信息

Onco Targets Ther. 2019 Jun 19;12:4729-4740. doi: 10.2147/OTT.S202938. eCollection 2019.

Abstract

Renal cell carcinoma (RCC) is one of the most common types of urological malignant tumors. Despite recent advances in diagnosis and management of RCC, its prognosis remains poor. Emerging evidence has shown that long noncoding RNAs (lncRNAs) play crucial regulatory roles in cancer biology. The most abundant transcript of long intergenic non-protein coding RNA p53 induced transcript () in clear cell RCC (ccRCC) was determined by RT-PCR. Quantitative real-time PCR was performed to examine expression in paired ccRCC samples and cell lines. The relationship of expression with clinicopathologic characteristics and clinical outcome was analyzed. The biological function of was studied by MTS and colony formation. The flow cytometry was used to analyze cell cycle distribution and apoptosis. The subcelluar fractionation and RIP assay was performed to explore the molecular mechanism of Western blotting and immunofluorescence was carried out to examine EZH2 and p53. We found that the was frequently upregulated in ccRCC samples. Furthermore, we observed that the level of depended on gender as well as on pT and TNM stage of patients with ccRCC. Moreover, patients with high expression had poor disease-free survival and overall survival. Functionally, overexpression of promoted ccRCC cell proliferation, induced cell cycle progression, and inhibited apoptosis. was primarily located in cell nuclei and interacted with EZH2. When EZH2 was knocked down in 769P and OS-RC-2 cells overexpressing , the effect of on cell proliferation, cell cycle, and apoptosis was partially reversed. Additionally, inducing p53 by doxorubicin (Dox) promoted expression. Collectively, our results provide novel insights into the important role of in ccRCC development and indicate that may serve as a valuable prognostic biomarker for ccRCC.

摘要

肾细胞癌(RCC)是泌尿系统最常见的恶性肿瘤类型之一。尽管近年来RCC的诊断和治疗取得了进展,但其预后仍然很差。新出现的证据表明,长链非编码RNA(lncRNAs)在癌症生物学中发挥着关键的调节作用。通过逆转录聚合酶链反应(RT-PCR)确定了透明细胞肾细胞癌(ccRCC)中长链基因间非蛋白质编码RNA p53诱导转录本()的最丰富转录本。进行定量实时聚合酶链反应以检测配对的ccRCC样本和细胞系中的表达。分析了表达与临床病理特征和临床结局的关系。通过MTS和集落形成研究了的生物学功能。采用流式细胞术分析细胞周期分布和凋亡情况。进行亚细胞分级分离和RNA免疫沉淀(RIP)试验以探索的分子机制,进行蛋白质免疫印迹法和免疫荧光检测以检测EZH2和p53。我们发现,在ccRCC样本中经常上调。此外,我们观察到的水平取决于性别以及ccRCC患者的pT和TNM分期。此外,高表达的患者无病生存期和总生存期较差。在功能上,的过表达促进了ccRCC细胞增殖,诱导了细胞周期进程,并抑制了凋亡。主要位于细胞核中并与EZH2相互作用。当在过表达的769P和OS-RC-2细胞中敲低EZH2时,对细胞增殖、细胞周期和凋亡的影响部分被逆转。此外,阿霉素(Dox)诱导p53可促进表达。总的来说,我们的结果为在ccRCC发展中的重要作用提供了新的见解,并表明可能作为ccRCC的一个有价值的预后生物标志物。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b11d/6592040/d55e7058cf13/OTT-12-4729-g0001.jpg

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