Novel circular RNA hsa_circ_0036683 suppresses proliferation and migration by mediating the miR-4664-3p/CDK2AP2 axis in non-small cell lung cancer.

IF 2.3 3区医学 Q3 ONCOLOGY Thoracic Cancer Pub Date : 2024-09-01 Epub Date: 2024-08-07 DOI:10.1111/1759-7714.15396

Rui Liu, Han Zhang, Jiaxuan Xin, Shu-Yang Xie, Fei Jiao, You-Jie Li, Meng-Yuan Chu, Junming Qiu, Yun-Fei Yan

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Abstract

Background: The aim of the present study was to investigate the function of novel circular RNA hsa_circ_0036683 (circ-36683) in non-small cell lung cancer (NSCLC).

Methods: RNA sequencing was used to screen out differentially expressed miRNAs. Expression levels of miR-4664-3p and circ-36683 were evaluated in lung carcinoma cells and tissues by quantitative reverse transcription-polymerase chain reaction (qRT-PCR). The effects of miR-4664-3p and circ-36683 on proliferation and migration were assessed using cell counting kit-8 (CCK-8), wound healing and transwell migration assays and xenograft experiments. The targeting relationship of circ-36683/miR-4664-3p/CDK2AP2 was assessed by luciferase reporter assays, western blot, qRT-PCR and argonaute2-RNA immunoprecipitation (AGO2 RIP). Co-immunoprecipitation (Co-IP), 5-ethynyl-2'-deoxyuridine (EdU) staining and CCK-8 were used to validate the indispensable role of CDK2AP2 in suppressing cell proliferation as a result of CDK2AP1 overexpression.

Results: By RNA sequencing, miR-4664-3p was screened out as an abnormally elevated miRNA in NSCLC tissues. Transfection of miR-4664-3p could promote cell proliferation, migration and xenograft tumor growth. As a target of miR-4664-3p, CDK2AP2 expression was downregulated by miR-4664-3p transfection and CDK2AP2 overexpression could abolish the proliferation promotion resulting from miR-4664-3p elevation. Circ-36683, derived from back splicing of ABHD2 pre-mRNA, was attenuated in NSCLC tissue and identified as a sponge of miR-4664-3p. The functional study revealed that circ-36683 overexpression suppressed cell proliferation, migration and resulted in G0/G1 phase arrest. More importantly, the antioncogenic function of circ-36683 was largely dependent on the miR-4664-3p/CDK2AP2 axis, through which circ-36683 could upregulate the expression of p53/p21/p27 and downregulate the expression of CDK2/cyclin E1.

Conclusion: The present study revealed the antioncogenic role of circ-36683 in suppressing cell proliferation and migration and highlighted that targeting the circ-36683/miR-4664-3p/CDK2AP2 axis is a promising strategy for the intervention of NSCLC.

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新型环状 RNA hsa_circ_0036683 通过介导 miR-4664-3p/CDK2AP2 轴抑制非小细胞肺癌的增殖和迁移。

研究背景本研究旨在探讨新型环状 RNA hsa_circ_0036683（circ-36683）在非小细胞肺癌（NSCLC）中的功能：方法：采用 RNA 测序筛选出差异表达的 miRNA。方法：采用 RNA 测序筛选出差异表达的 miRNA，并通过反转录聚合酶链反应（qRT-PCR）定量评估 miR-4664-3p 和 circ-36683 在肺癌细胞和组织中的表达水平。使用细胞计数试剂盒-8（CCK-8）、伤口愈合和跨孔迁移试验以及异种移植实验评估了 miR-4664-3p 和 circ-36683 对增殖和迁移的影响。通过荧光素酶报告实验、Western 印迹、qRT-PCR 和 argonaute2-RNA 免疫沉淀（AGO2 RIP）评估了 circ-36683/miR-4664-3p/CDK2AP2 的靶向关系。共免疫沉淀（Co-IP）、5-乙炔基-2'-脱氧尿苷（EdU）染色和 CCK-8 被用来验证 CDK2AP2 在 CDK2AP1 过表达抑制细胞增殖中不可或缺的作用：结果：通过RNA测序筛选出miR-4664-3p是NSCLC组织中异常升高的miRNA。转染 miR-4664-3p 可促进细胞增殖、迁移和异种移植肿瘤的生长。作为miR-4664-3p的靶标，CDK2AP2的表达因miR-4664-3p转染而下调，CDK2AP2的过表达可消除miR-4664-3p升高对细胞增殖的促进作用。Circ-36683来自于ABHD2前mRNA的反向剪接，在NSCLC组织中被减弱，并被鉴定为miR-4664-3p的海绵。功能研究显示，circ-36683 的过表达抑制了细胞的增殖和迁移，并导致 G0/G1 期停滞。更重要的是，circ-36683的抗诱导功能主要依赖于miR-4664-3p/CDK2AP2轴，通过该轴，circ-36683可以上调p53/p21/p27的表达，下调CDK2/细胞周期蛋白E1的表达：本研究揭示了circ-36683在抑制细胞增殖和迁移中的抗原性作用，并强调靶向circ-36683/miR-4664-3p/CDK2AP2轴是干预NSCLC的一种有前景的策略。

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

Thoracic Cancer ONCOLOGY-RESPIRATORY SYSTEM

CiteScore

5.20

自引率

3.40%

发文量

439

审稿时长

2 months

期刊介绍： Thoracic Cancer aims to facilitate international collaboration and exchange of comprehensive and cutting-edge information on basic, translational, and applied clinical research in lung cancer, esophageal cancer, mediastinal cancer, breast cancer and other thoracic malignancies. Prevention, treatment and research relevant to Asia-Pacific is a focus area, but submissions from all regions are welcomed. The editors encourage contributions relevant to prevention, general thoracic surgery, medical oncology, radiology, radiation medicine, pathology, basic cancer research, as well as epidemiological and translational studies in thoracic cancer. Thoracic Cancer is the official publication of the Chinese Society of Lung Cancer, International Chinese Society of Thoracic Surgery and is endorsed by the Korean Association for the Study of Lung Cancer and the Hong Kong Cancer Therapy Society. The Journal publishes a range of article types including: Editorials, Invited Reviews, Mini Reviews, Original Articles, Clinical Guidelines, Technological Notes, Imaging in thoracic cancer, Meeting Reports, Case Reports, Letters to the Editor, Commentaries, and Brief Reports.