RGS1 stabilized by METTL3-mediated m6A modification promotes the tumorigenicity and macrophage M2 polarization in osteosarcoma

IF 3.5 2区医学 Q2 Medicine

Journal of Bone Oncology Pub Date : 2025-07-26 DOI:10.1016/j.jbo.2025.100705

Zhizhong Liang , Yuxia Shi , Mao Wang , Liqiang Zhang

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Abstract

Background

Regulators of G-protein signaling 1 (RGS1) has been reported to be involved in immune cell regulation in many cancer types. However, the specific role and mechanism in osteosarcoma (OS) progression and macrophage activation remain unclear.

Methods

Levels of mRNA and protein were examined using qRT-PCR and western blotting. Transwell assay, wound healing assay, EdU assay and flow cytometry were used to investigate OS cell invasion, migration, proliferation and apoptosis. Xenografts in mice were established for in vivo assay. Macrophage M2 polarization was evaluated by detecting CD206 + macrophages by flow cytometry. ELISA analysis detected IL-6 and TGF-β1 levels. Methylated RNA immunoprecipitation assay was applied to explore the specific binding of RGS1 and METTL3 (methyltransferase-like 3).

Results

RGS1 was highly expressed in OS tissues and cells. The silencing of RGS1 suppressed OS cell invasion, migration, growth and impaired immune response by inhibiting macrophage M2 polarization and M2 macrophage-mediated release of IL-10 and TGF-β1. Mechanistically, METTL3 promoted RGS1 m6A modification and stabilized its expression. METTL3 deficiency also inhibited OS cell invasion, migration, growth and macrophage M2 polarization, while these effects could be abolished by RGS1 overexpression. Besides that, IL–10 elevation induced by M2 macrophages promoted OS cell oncogenic phenotypes.

Conclusion

METTL3 stabilized RGS1 mRNA in an m6A-dependent manner to promote the tumorigenicity and macrophage M2 polarization in osteosarcoma, suggesting a novel insight into the therapy of osteosarcoma.

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经mettl3介导的m6A修饰稳定的RGS1促进骨肉瘤的致瘤性和巨噬细胞M2极化

据报道，g蛋白信号1 （RGS1）的调节因子参与许多癌症类型的免疫细胞调节。然而，在骨肉瘤（OS）进展和巨噬细胞活化中的具体作用和机制尚不清楚。方法采用qRT-PCR和western blotting检测mRNA和蛋白水平。采用Transwell法、创面愈合法、EdU法和流式细胞术观察骨肉瘤细胞的侵袭、迁移、增殖和凋亡情况。建立小鼠异种移植物进行体内实验。流式细胞术检测CD206 +巨噬细胞，评价巨噬细胞M2极化。ELISA法检测IL-6、TGF-β1水平。采用甲基化RNA免疫沉淀法研究RGS1与甲基转移酶样3 （methyltransferase-like 3）的特异性结合。结果rgs1在骨肉瘤组织和细胞中高表达。RGS1的沉默通过抑制巨噬细胞M2极化和M2巨噬细胞介导的IL-10和TGF-β1的释放来抑制OS细胞的侵袭、迁移、生长和免疫应答受损。机制上，METTL3促进了RGS1 m6A的修饰并稳定了其表达。METTL3缺乏还能抑制OS细胞的侵袭、迁移、生长和巨噬细胞M2极化，而这些作用可通过RGS1过表达而消除。此外，M2巨噬细胞诱导IL-10升高可促进OS细胞的致癌表型。结论mettl3以m6a依赖的方式稳定RGS1 mRNA，促进骨肉瘤的致瘤性和巨噬细胞M2极化，为骨肉瘤的治疗提供了新的思路。

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来源期刊

Journal of Bone Oncology ONCOLOGY-

CiteScore

7.20

自引率

2.90%

发文量

审稿时长

34 days

期刊介绍： The Journal of Bone Oncology is a peer-reviewed international journal aimed at presenting basic, translational and clinical high-quality research related to bone and cancer. As the first journal dedicated to cancer induced bone diseases, JBO welcomes original research articles, review articles, editorials and opinion pieces. Case reports will only be considered in exceptional circumstances and only when accompanied by a comprehensive review of the subject. The areas covered by the journal include: Bone metastases (pathophysiology, epidemiology, diagnostics, clinical features, prevention, treatment) Preclinical models of metastasis Bone microenvironment in cancer (stem cell, bone cell and cancer interactions) Bone targeted therapy (pharmacology, therapeutic targets, drug development, clinical trials, side-effects, outcome research, health economics) Cancer treatment induced bone loss (epidemiology, pathophysiology, prevention and management) Bone imaging (clinical and animal, skeletal interventional radiology) Bone biomarkers (clinical and translational applications) Radiotherapy and radio-isotopes Skeletal complications Bone pain (mechanisms and management) Orthopaedic cancer surgery Primary bone tumours Clinical guidelines Multidisciplinary care Keywords: bisphosphonate, bone, breast cancer, cancer, CTIBL, denosumab, metastasis, myeloma, osteoblast, osteoclast, osteooncology, osteo-oncology, prostate cancer, skeleton, tumour.