据报道，癌症相关成纤维细胞（CAF）可以影响癌细胞增殖、转移、铁死亡和免疫逃逸。 METTL3 介导的 N6-甲基腺嘌呤 (m6A) 修饰参与结直肠癌 (CRC) 的肿瘤发生。在此，我们研究了 CAF 衍生的外泌体 (exo) 中 METTL3 依赖性 m6A 是否影响 CRC 进展。qRT-PCR 和蛋白质印迹分析检测了 mRNA 和蛋白质的水平。分别使用 MTT、集落形成、Transwell 和伤口愈合测定来评估细胞增殖和转移。通过检测细胞活力以及erastin处理后Fe、活性氧和谷胱甘肽的水平来评估细胞铁死亡。通过超速离心从 CAF 中分离出外泌体。通过甲基化 RNA 免疫沉淀测定确定 m6A 修饰谱，并使用双荧光素酶报告基因测定验证 METTL3 和 ACSL3（酰基辅酶 A 合成酶 3）之间的相互作用。建立动物模型进行体内分析。CAFs促进CRC细胞增殖和转移，抑制细胞铁死亡。 METTL3 在 CAF 中富集并被包装到外泌体中。 CRC 样本中 m6A 修饰和 METTL3 表达增加。 CAFs-exo 中 METTL3 的敲低抑制了 CRC 细胞增殖和转移，并诱导细胞铁死亡。从机制上讲，METTL3 诱导 ACSL3 m6A 修饰并稳定其表达。 METTL3 沉默的 CAFs-exo 介导的抗癌作用可以通过 ACSL3 过表达来挽救。此外，体内实验还表明，METTL3减少的CAFs-exo可以阻碍小鼠模型中的CRC生长和转移。CAFs通过外泌体METTL3引发的ACSL3 m6A修饰促进增殖和转移，并抑制CRC的铁死亡。© 2024。作者。

Cancer-associated fibroblasts (CAFs) have been reported that can affect cancer cell proliferation, metastasis, ferroptosis, and immune escape. METTL3-mediated N6-methyladenine (m6A) modification is involved in the tumorigenesis of colorectal cancer (CRC). Herein, we investigated whether METTL3-dependent m6A in CAFs-derived exosomes (exo) affected CRC progression.qRT-PCR and western blotting analyses detected levels of mRNAs and proteins. Cell proliferation and metastasis were evaluated using MTT, colony formation, transwell, and wound healing assays, respectively. Cell ferroptosis was assessed by detecting cell viability and the levels of Fe+, reactive oxygen species, and glutathione after erastin treatment. Exosomes were isolated from CAFs by ultracentrifugation. The m6A modification profile was determined by methylated RNA immunoprecipitation assay and the interaction between METTL3 and ACSL3 (acyl-CoA synthetase 3) was verified using dual-luciferase reporter assay. Animal models were established for in vivo analysis.CAFs promoted CRC cell proliferation and metastasis, and suppressed cell ferroptosis. METTL3 was enriched in CAFs and was packaged into exosomes. The m6A modification and METTL3 expression were increased in CRC samples. Knockdown of METTL3 in CAFs-exo suppressed CRC cell proliferation and metastasis, and induced cell ferroptosis. Mechanistically, METTL3 induced ACSL3 m6A modification and stabilized its expression. The anticancer effects mediated by METTL3-silenced CAFs-exo could be rescued by ACSL3 overexpression. Moreover, in vivo assay also showed that CAFs-exo with decreased METTL3 could hinder CRC growth and metastasis in mice models.CAFs promoted the proliferation and metastasis, and restrained the ferroptosis in CRC by exosomal METTL3-elicited ACSL3 m6A modification.© 2024. The Author(s).