METTL3 promotes the progression of osteosarcoma through the N6-methyladenosine modification of MCAM via IGF2BP1.

IF 5.7 2区生物学 Q1 BIOLOGY Biology Direct Pub Date : 2024-06-07 DOI:10.1186/s13062-024-00486-x

Dongjian Song, Qi Wang, Zechen Yan, Meng Su, Hui Zhang, Longyan Shi, Yingzhong Fan, Qian Zhang, Heying Yang, Da Zhang, Qiuliang Liu

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Abstract

Background: The molecular mechanisms of osteosarcoma (OS) are complex. In this study, we focused on the functions of melanoma cell adhesion molecule (MCAM), methyltransferase 3 (METTL3) and insulin like growth factor 2 mRNA binding protein 1 (IGF2BP1) in OS development.

Methods: qRT-PCR assay and western blot assay were performed to determine mRNA and protein expression of MCAM, METTL3, IGF2BP1 and YY1. MTT assay and colony formation assay were conducted to assess cell proliferation. Cell apoptosis, invasion and migration were evaluated by flow cytometry analysis, transwell assay and wound-healing assay, respectively. Methylated RNA Immunoprecipitation (MeRIP), dual-luciferase reporter, Co-IP, RIP and ChIP assays were performed to analyze the relationships of MCAM, METTL3, IGF2BP1 and YY1. The functions of METTL3 and MCAM in tumor growth were explored through in vivo experiments.

Results: MCAM was upregulated in OS, and MCAM overexpression promoted OS cell growth, invasion and migration and inhibited apoptosis. METTL3 and IGF2BP1 were demonstrated to mediate the m6A methylation of MCAM. Functionally, METTL3 or IGF2BP1 silencing inhibited OS cell progression, while MCAM overexpression ameliorated the effects. Transcription factor YY1 promoted the transcription level of METTL3 and regulated METTL3 expression in OS cells. Additionally, METTL3 deficiency suppressed tumor growth in vivo, while MCAM overexpression abated the effect.

Conclusion: YY1/METTL3/IGF2BP1/MCAM axis aggravated OS development, which might provide novel therapy targets for OS.

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METTL3通过IGF2BP1对MCAM进行N6-甲基腺苷修饰，从而促进骨肉瘤的进展。

背景：骨肉瘤（OS）的分子机制十分复杂。方法：通过 qRT-PCR 检测和 Western 印迹检测确定 MCAM、METTL3、IGF2BP1 和 YY1 的 mRNA 和蛋白表达。MTT 试验和集落形成试验用于评估细胞增殖。细胞凋亡、侵袭和迁移分别通过流式细胞仪分析、Transwell 试验和伤口愈合试验进行评估。通过甲基化 RNA 免疫沉淀（MeRIP）、双荧光素酶报告、Co-IP、RIP 和 ChIP 检测分析 MCAM、METTL3、IGF2BP1 和 YY1 的关系。通过体内实验探讨了METTL3和MCAM在肿瘤生长中的功能：结果：MCAM在OS中上调，MCAM过表达促进OS细胞的生长、侵袭和迁移，并抑制细胞凋亡。METTL3和IGF2BP1被证实介导了MCAM的m6A甲基化。在功能上，沉默METTL3或IGF2BP1可抑制OS细胞的进展，而过表达MCAM则可改善其效果。转录因子YY1可促进METTL3的转录水平，并调节METTL3在OS细胞中的表达。此外，METTL3缺乏可抑制肿瘤在体内的生长，而MCAM过表达则可减轻这种影响：结论：YY1/METTL3/IGF2BP1/MCAM轴加重了OS的发展，可能为OS提供了新的治疗靶点。

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

Biology Direct 生物-生物学

CiteScore

6.40

自引率

10.90%

发文量

审稿时长

7 months

期刊介绍： Biology Direct serves the life science research community as an open access, peer-reviewed online journal, providing authors and readers with an alternative to the traditional model of peer review. Biology Direct considers original research articles, hypotheses, comments, discovery notes and reviews in subject areas currently identified as those most conducive to the open review approach, primarily those with a significant non-experimental component.