环状 RNA 血管生成素相关蛋白通过 miR-876-5p/WNT5A 轴发挥致癌作用在非小细胞肺癌中。

CircVAPA exerts oncogenic property in non-small cell lung cancer by the miR-876-5p/WNT5A axis.

机构信息

Department of Medical Oncology, Huazhong University of Science and Technology Union Shenzhen Hospital, Shenzhen, China.

Department of Medical Oncology, The First Affiliated Hospital, Sun Yat-Sen University, Guangzhou, Guangdong, China.

出版信息

J Gene Med. 2021 Jun;23(6):e3325. doi: 10.1002/jgm.3325. Epub 2021 Apr 12.

DOI:10.1002/jgm.3325

PMID:33619796

Abstract

BACKGROUND

Non-small cell lung cancer (NSCLC) is one of the most fatal malignant tumors. Emerging studies have clarified the crucial roles of circular RNAs (circRNAs) in the tumorigenesis of cancers. CircVAPA was demonstrated to function in some human cancers. The present study aimed to investigate the role of circVAPA in NSCLC.

METHODS

A quantitative real-time polymerase chain reaction was used to measure the expression of genes. Actinomycin D and RNase R were employed to examine the stability of circVAPA. Cell-counting kit-8, 5-ethynyl-2'-deoxyuridine, Transwell and sphere formation assays, and well as western blot analysis, were conducted to examine the changes of NSCLC cells in response to circVAPA knockdown. A luciferase reporter assay was conducted for the molecular mechanism.

RESULTS

Our findings demonstrated high expression of circVAPA in tissues and cell lines of NSCLC. Knockdown of circVAPA had a suppressive effect on cell proliferation, migration, invasion and stemness, and also inhibited tumor growth in vivo. Mechanistically, circVAPA acted as a competing endogenous RNA to up-regulate WNT5A by sponging miR-876-5p. Moreover, circVAPA activated Wnt/β-catenin signaling by up-regulation of WNT5A. Rescue assays showed that silencing of miR-876-5p or overexpression of WNT5A reversed the circVAPA knockdown-mediated inhibition on cellular processes in NSCLC.

CONCLUSIONS

CircVAPA promotes aggressive phenotypes of NSCLC cells by the miR-876-5p/WNT5A axis activating Wnt/β-catenin signaling.

摘要

背景

非小细胞肺癌（NSCLC）是最致命的恶性肿瘤之一。新兴研究已经阐明了环状 RNA（circRNA）在癌症发生中的关键作用。CircVAPA 被证明在一些人类癌症中发挥作用。本研究旨在探讨 circVAPA 在 NSCLC 中的作用。

方法

采用实时定量聚合酶链反应测定基因表达。采用放线菌素 D 和 RNase R 检测 circVAPA 的稳定性。细胞计数试剂盒-8、5-乙炔基-2'-脱氧尿苷、Transwell 和球体形成试验以及 Western blot 分析用于检测 circVAPA 敲低后 NSCLC 细胞的变化。采用荧光素酶报告基因检测进行分子机制研究。

结果

我们的研究结果表明，circVAPA 在 NSCLC 组织和细胞系中表达较高。circVAPA 敲低对细胞增殖、迁移、侵袭和干性有抑制作用，并抑制体内肿瘤生长。机制上，circVAPA 作为竞争性内源性 RNA 通过海绵吸附 miR-876-5p 而上调 WNT5A。此外，circVAPA 通过上调 WNT5A 激活 Wnt/β-catenin 信号通路。挽救实验表明，沉默 miR-876-5p 或过表达 WNT5A 逆转了 circVAPA 敲低介导的 NSCLC 细胞中细胞过程的抑制。