N⁶-methyladenosine (m⁶A) is the most universal internal RNA modification on messenger RNAs and regulates the fate and functions of m⁶A-modified transcripts through m⁶A-specific binding proteins. Nevertheless, the functional role and potential mechanism of the m⁶A reading proteins in ocular melanoma tumorigenicity, especially cancer stem-like cell (CSC) properties, remain to be elucidated. Herein, we demonstrated that the m⁶A reading protein YTHDF3 promotes the translation of the target transcript CTNNB1, contributing to ocular melanoma propagation and migration through m⁶A methylation. YTHDF3 is highly expressed in ocular melanoma stem-like cells and abundantly enriched in ocular melanoma tissues, which is related to poor clinical prognosis. Moreover, YTHDF3 is required for the maintenance of CSC properties and tumor initiation capacity in ocular melanoma both in vitro and in vivo. Ocular melanoma cells with targeted YTHDF3 knockdown exhibited inhibitory tumor proliferation and migration abilities. Transcriptome-wide mapping of m⁶A peaks and YTHDF3 binding peaks on mRNAs revealed a key target gene candidate, CTNNB1. Mechanistically, YTHDF3 enhances CTNNB1 translation through recognizing and binding the m⁶A peaks on CTNNB1 mRNA.

N ⁶ -甲基腺苷 (m ⁶ A) 是对信使 RNA 最普遍的内部 RNA 修饰，并通过 m 6 A 特异性结合蛋白调节 m 6 A 修饰转录物的^命运和^功能。然而，m ⁶ A 读取蛋白在眼黑色素瘤致瘤性中的功能作用和潜在机制，尤其是癌症干细胞样细胞 (CSC) 特性，仍有待阐明。在此，我们证明了 m ^{6 A 阅读蛋白 YTHDF3 促进了目标转录物}CTNNB1的翻译，通过 m ⁶ A 甲基化促进了眼黑色素瘤的传播和迁移。YTHDF3在眼部黑色素瘤干细胞样细胞中高表达，并在眼部黑色素瘤组织中大量富集，与临床预后不良有关。此外，YTHDF3是在体外和体内维持眼部黑色素瘤的 CSC 特性和肿瘤起始能力所必需的。具有靶向YTHDF3敲低的眼黑色素瘤细胞表现出抑制性肿瘤增殖和迁移能力。^{m 6} A 峰和 YTHDF3 结合峰在 mRNA 上的全转录组作图揭示了关键靶基因候选CTNNB1。从机制上讲，YTHDF3通过识别和结合CTNNB1 mRNA 上的m ⁶ A 峰来增强CTNNB1翻译。