MicroRNA-501-3p inhibits the proliferation of kidney cancer cells by targeting WTAP.
MicroRNA-501-3p inhibits the proliferation of kidney cancer cells by targeting WTAP.
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MicroRNA-501-3p 通过靶向 WTAP 抑制肾癌细胞的增殖。
DOI：
10.1002/cam4.4157
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发表时间：
2021-10
期刊：
影响因子：
4
通讯作者：
Xie L
中科院分区：
文献类型：
--
作者：
He L;Chen S;Ying Y;Xie H;Li J;Ma X;Wang W;Shen H;Wang X;Zheng X;Xie L
越来越多的证据表明miR-501-3p在多种肿瘤的发生发展中起重要作用。然而，它在肾癌(RCC)中的作用和潜在机制仍有待阐明。应用定量RT-PCR、Western印迹和生物信息学方法检测miR-501-3p和Wilms肿瘤相关蛋白(WTAP)在肾癌细胞系和临床组织中的表达。采用流式细胞仪、集落形成和CCK8实验观察miR-501-3p对肾癌细胞增殖的影响。经Western blotting、qRT-PCR和双荧光素酶报告基因鉴定，证实miR-501-3p为目的基因。用斑点印迹法检测miR-501-3p过表达或敲除后N6-甲基腺苷(M6A)的RNA甲基化水平。MIR-501-3p在人肾细胞癌细胞系和组织中的表达显著下调。相反，它的过表达通过诱导G1期阻滞而显著抑制体外癌细胞的增殖。此外，WTAP基因被证实为miR-501-3p的直接靶基因。WTAP基因敲除单独有效地产生了与miR-501-3p过表达相同的癌症抑制效果，在这两种情况下，肾癌细胞中m6A的水平都降低了。救援实验进一步证实了miR-501-3P与WTAP之间的分子间相互作用。肾癌的进展受miR-501-3p/WTAP/CDK2轴的调控，并受miR-501-3p过表达的抑制。MIR-501-3p在人肾癌细胞株和组织中的表达显著下调。此外，WTAP基因被确认为miR-501-3p的直接靶基因。肾癌的进展是通过miR-501-3p/WTAP/CDK2轴调节的，并受到microRNA过表达的抑制。
Emerging evidence suggests that miR‐501‐3p plays an important role in the pathogenesis and progression of various carcinomas. However, its role and underlying mechanisms in renal cell carcinoma (RCC) remain to be elucidated. Quantitative RT‐PCR, western blot, and bioinformatics methods were used to evaluate the expression of miR‐501‐3p and Wilms’ tumor 1‐associating protein (WTAP) in RCC cell lines and clinical tissues. The effects of miR‐501‐3p on the proliferation of RCC cells were investigated using flow cytometric, colony formation, and CCK8 assays. The target gene of miR‐501‐3p was confirmed by western blotting, qRT‐PCR, and dual‐luciferase reporter assays. The levels of RNA methylation with N6‐methyladenosine (m6A) following miR‐501‐3p overexpression or knockdown of its target gene were quantified using a dot‐blot assay. miR‐501‐3p expression was significantly downregulated in human RCC cell lines and tissues. In contrast, its overexpression markedly inhibited cancer cell proliferation in vitro by inducing G1 phase arrest. Moreover, WTAP was verified as a direct target gene of miR‐501‐3p. WTAP gene knockdown alone efficiently produced the same cancer‐inhibiting effects as miR‐501‐3p overexpression, with the level of m6A in RCC cells being decreased under both scenarios. The intermolecular interaction between miR‐501‐3p and WTAP was further substantiated by rescue experiments. RCC progression is regulated via the miR‐501‐3p/WTAP/CDK2 axis and is inhibited by the overexpression of miR‐501‐3p. miR‐501‐3p was significantly down‐regulated in human RCC cell lines and tissues. Moreover, WTAP was confirmed as the direct target gene of miR‐501‐3p. RCC progression is regulated via the miR‐501‐3p/WTAP/CDK2 axis and is inhibited by the overexpression of the microRNA.
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