NOP2-mediated 5-methylcytosine modification of APOL1 messenger RNA activates PI3K-Akt and facilitates clear cell renal cell carcinoma progression

IF 8.2 2区 生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY
Junjie Tian, Jianguo Gao, Cheng Cheng, Zhijie Xu, Xiaoyi Chen, Yunfei Wu, Guanghou Fu, Baiye Jin
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引用次数: 0

Abstract

Background: By regulating the functions of multiple RNAs, 5-methylcytosine (m5C) RNA methylation, particularly mediated by NOP2, is involved in tumorigenesis and developments. However, the specific functions and potential mechanisms of m5C, especially involving NOP2, in clear-cell renal cell carcinoma (ccRCC), remain unclear./nMethods: NOP2 expression in cell lines and patient tissues was detected using western blotting, quantitative real-time polymerase chain reaction (RT-qPCR), and immunohistochemistry. The biological effects of NOP2 on ccRCC cells were investigated through a series of in vitro and in vivo experiments. To explore the potential regulatory mechanisms by which NOP2 affects ccRCC progression, m5C bisulfite sequencing, RNA-sequencing, RNA immunoprecipitation and methylated RNA immunoprecipitation (RIP/MeRIP) RT-qPCR assay, luciferase reporter assay, RNA stability assay, and bioinformatic analysis were performed./nResults: NOP2 expression was significantly upregulated in ccRCC tissues and was associated with poor prognosis. Moreover, loss-of-function and gain-of-function assays demonstrated that NOP2 altered ccRCC cell proliferation, migration, and invasion. Mechanistically, NOP2 stimulated m5C modification of apolipoprotein L1 (APOL1) mRNA, and m5C reader YBX1 stabilized APOL1 mRNA through recognizing and binding to m5C site in the 3′-untranslated regions. Silencing APOL1 expression inhibited ccRCC cell proliferation in vitro and tumor formation in vivo. Furthermore, NOP2/APOL1 affected ccRCC progression via the PI3K-Akt signaling pathway./nConclusion: NOP2 functions as an oncogene in ccRCC by promoting tumor progression through the m5C-dependent stabilization of APOL1, which in turn regulates the PI3K-Akt signaling pathway, suggesting a potential therapeutic target for ccRCC.
NOP2 介导的 APOL1 信使 RNA 的 5-甲基胞嘧啶修饰可激活 PI3K-Akt 并促进透明细胞肾细胞癌的进展
背景:通过调节多种 RNA 的功能,5-甲基胞嘧啶(m5C)RNA 甲基化,尤其是由 NOP2 介导的甲基化,参与了肿瘤的发生和发展。然而,m5C(尤其是 NOP2)在透明细胞肾细胞癌(ccRCC)中的具体功能和潜在机制仍不清楚:采用免疫印迹、实时定量聚合酶链反应(RT-qPCR)和免疫组织化学方法检测细胞系和患者组织中 NOP2 的表达。通过一系列体外和体内实验研究了 NOP2 对 ccRCC 细胞的生物学效应。为了探索NOP2影响ccRCC进展的潜在调控机制,研究人员进行了m5C亚硫酸氢盐测序、RNA测序、RNA免疫沉淀和甲基化RNA免疫沉淀(RIP/MeRIP)RT-qPCR检测、荧光素酶报告检测、RNA稳定性检测和生物信息学分析:NOP2在ccRCC组织中表达明显上调,并与不良预后相关。此外,功能缺失和功能增益实验表明,NOP2 可改变 ccRCC 细胞的增殖、迁移和侵袭。从机理上讲,NOP2刺激脂蛋白L1(APOL1)mRNA的m5C修饰,m5C阅读器YBX1通过识别并结合3′-非翻译区的m5C位点稳定APOL1 mRNA。抑制 APOL1 的表达可抑制体外 ccRCC 细胞的增殖和体内肿瘤的形成。此外,NOP2/APOL1通过PI3K-Akt信号通路影响ccRCC的进展:NOP2通过m5C依赖性稳定APOL1,进而调节PI3K-Akt信号通路,促进肿瘤进展,从而在ccRCC中发挥癌基因的作用,为ccRCC提供了潜在的治疗靶点。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
International Journal of Biological Sciences
International Journal of Biological Sciences 生物-生化与分子生物学
CiteScore
16.90
自引率
1.10%
发文量
413
审稿时长
1 months
期刊介绍: The International Journal of Biological Sciences is a peer-reviewed, open-access scientific journal published by Ivyspring International Publisher. It dedicates itself to publishing original articles, reviews, and short research communications across all domains of biological sciences.
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