METTL3/IGF2BP1 influences the development of non-small-cell lung cancer by mediating m6A methylation modification of TRPV1.

IF 2.3 3区医学 Q3 ONCOLOGY Thoracic Cancer Pub Date : 2024-09-01 Epub Date: 2024-08-01 DOI:10.1111/1759-7714.15381

Wenjie Bai, Gang Xiao, Guijing Xie, Zhibo Chen, Xie Xu, Jie Zeng, Jianjiang Xie

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Abstract

Background: Methyltransferase 3 (METTL3) accelerates N6-methyladenosine (m6A) modifications and affects cancer progression, including non-small-cell lung cancer (NSCLC). In this study, we aimed to explore the regulatory mechanisms of METTL3 underling NSCLC.

Methods: Immunohistochemical assay, quantitative real-time polymerase chain reaction (qRT-PCR) assay, and western blot assay were conducted for gene expression. MTT assay and colony formation assay were performed to explore cell proliferation capacity. Cell apoptosis and THP-1 cell polarization were estimated by flow cytometry analysis. Cell migration and invasion capacities were evaluated by transwell assay. Methylated RNA immunoprecipitation assay, dual-luciferase reporter assay, actinomycin D treatment and RIP assay were performed to analyze the relationships of METTL3, insulin-like growth factor 2 mRNA binding protein 1 (IGF2BP1), and transient receptor potential cation channel subfamily V member 1 (TRPV1). The functions of METTL3 and TRPV1 in vivo were investigated through establishing the murine xenograft model.

Results: TRPV1 expression was upregulated in NSCLC and related poor prognosis. TRPV1 silencing inhibited NSCLC cell growth and metastasis, induced NSCLC cell apoptosis, and repressed M2 macrophage polarization. The results showed that METTL3 and IGF2BP1 could regulate TRPV1 expression through m6A methylation modification. Moreover, METTL3 deficiency inhibited NSCLC cell growth, metastasis, and M2 macrophage polarization and facilitated NSCLC cell apoptosis, while TRPV1 overexpression restored the impacts. In addition, METTL3 knockdown restrained tumor growth in vivo via regulating TRPV1 expression.

Conclusion: METTL3 bound to IGF2BP1 and enhanced IGF2BP1's m6A recognition of TRPV1 mRNA, thereby promoting NSCLC cell growth and metastasis, and inhibiting M2 macrophage polarization.

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METTL3/IGF2BP1通过介导TRPV1的m6A甲基化修饰影响非小细胞肺癌的发展。

背景：甲基转移酶3（METTL3）可加速N6-甲基腺苷（m6A）的修饰并影响癌症的进展，包括非小细胞肺癌（NSCLC）。本研究旨在探讨METTL3在NSCLC中的调控机制：方法：采用免疫组化、实时定量聚合酶链反应（qRT-PCR）和免疫印迹法检测基因表达。MTT 试验和集落形成试验用于检测细胞增殖能力。流式细胞术分析估计了细胞凋亡和 THP-1 细胞极化。细胞迁移和侵袭能力通过透孔试验进行评估。甲基化 RNA 免疫沉淀实验、双荧光素酶报告实验、放线菌素 D 处理和 RIP 实验分析了 METTL3、胰岛素样生长因子 2 mRNA 结合蛋白 1（IGF2BP1）和瞬时受体电位阳离子通道 V 亚家族成员 1（TRPV1）之间的关系。通过建立小鼠异种移植模型，研究了 METTL3 和 TRPV1 在体内的功能：结果：TRPV1在NSCLC中表达上调，与预后不良有关。沉默 TRPV1 可抑制 NSCLC 细胞生长和转移，诱导 NSCLC 细胞凋亡，抑制 M2 巨噬细胞极化。结果表明，METTL3和IGF2BP1可通过m6A甲基化修饰调控TRPV1的表达。此外，METTL3缺失可抑制NSCLC细胞生长、转移和M2巨噬细胞极化，促进NSCLC细胞凋亡，而TRPV1过表达则可恢复其影响。此外，METTL3敲除可通过调节TRPV1的表达抑制体内肿瘤的生长：结论：METTL3与IGF2BP1结合，增强了IGF2BP1对TRPV1 mRNA的m6A识别，从而促进了NSCLC细胞的生长和转移，抑制了M2巨噬细胞的极化。

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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.