YT521‐B homology (YTH) domain family (YTHDF) proteins serve as readers that directly recognise m6A modifications. In this study, we aim to probe the role of YTHDF1 in environmental carcinogen‐induced malignant transformation of gastric cells and gastric cancer (GC) carcinogenesis. We established a long‐term low‐dose MNU‐induced malignant transformation model in gastric epithelial cells. In vivo and in vitro experiments were conducted to validate the malignant phenotype and characterise the roles of YTHDF1 and its downstream genes in malignant transformation cells. Additionally, we explored downstream m6A modification targets of YTHDF1 using RNA‐sequencing, RNA immunoprecipitation, and proteomics analyses, and conducted validation experiments in cell experiments and clinical samples. Long‐term low‐dose exposure of MNU converted normal Gges‐1 cells into malignant cells. YTHDF1 mRNA and protein expression are increased in MNU‐induced malignant cells (p<0.001). Meanwhile, YTHDF1 knockdown inhibits the malignant potential of MNU‐treated cells (p<0.01). YTHDF1 knockdown specifically suppresses HSPH1 protein, but not RNA levels. RIP‐qPCR validates HSPH1 is the target of YTHDF1 (p<0.01). HSPH1 knockdown impairs the malignant potential of MNU‐induced transformed cells. The increased expression of the key regulatory factor YTHDF1 in MNU‐induced gastric carcinogenesis affects malignant transformation and tumorigenesis by regulating the translation of downstream HSPH1. These findings provide new potential targets for preventing and treating environmental chemical‐induced gastric carcinogenesis.

中文翻译：

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YTHDF1通过控制HSPH1翻译介导N-甲基-N-亚硝基脲诱导的胃癌发生

YT521-B 同源 (YTH) 结构域家族 (YTHDF) 蛋白充当直接识别 m6A 修饰的读取器。在本研究中，我们旨在探讨YTHDF1在环境致癌物诱导的胃细胞恶性转化和胃癌（GC）癌变中的作用。我们建立了长期低剂量 MNU 诱导胃上皮细胞恶性转化模型。进行体内和体外实验来验证恶性表型并表征 YTHDF1 及其下游基因在恶性转化细胞中的作用。此外，我们利用RNA测序、RNA免疫沉淀和蛋白质组学分析探索了YTHDF1的下游m6A​​修饰靶点，并在细胞实验和临床样本中进行了验证实验。长期低剂量暴露 MNU 将正常 Gges-1 细胞转化为恶性细胞。MNU 诱导的恶性细胞中 YTHDF1 mRNA 和蛋白表达增加（p<0.001）。同时，YTHDF1 敲低抑制了 MNU 处理细胞的恶性潜能（p<0.01）。YTHDF1 敲除特异性抑制 HSPH1 蛋白，但不抑制 RNA 水平。RIP-qPCR 验证 HSPH1 是 YTHDF1 的靶标（p<0.01）。HSPH1 敲低会损害 MNU 诱导的转化细胞的恶性潜能。MNU诱导的胃癌发生中关键调节因子YTHDF1表达增加，通过调节下游HSPH1的翻译影响恶性转化和肿瘤发生。这些发现为预防和治疗环境化学物质诱发的胃癌提供了新的潜在靶标。