N6-甲基腺苷 (m6A) 修饰在调节 RNA 成熟、稳定性和翻译中发挥着重要作用。因此，m6A 修饰参与多种病理生理过程，包括肝细胞癌 (HCC)。然而，m6A 修饰对 HCC 中 RNA 功能的直接贡献仍不清楚。在这里，我们将 LEAWBIH（在 HCC 中表观遗传激活 Wnt/β-catenin 信号传导的长非编码 RNA）鉴定为 m6A 修饰的长非编码 RNA (lncRNA)，并研究了 m6A 对 HCC 中 LEAWBIH 功能的影响。进行聚合酶链反应来测量组织和细胞中的基因表达。使用甲基化 RNA 免疫沉淀测定和基于单碱基延伸和连接的 qPCR 扩增方法检测 m6A 修饰水平。使用 Glo 细胞活力和 CCK-8 测定法评估细胞增殖。使用Transwell迁移和侵袭测定评估细胞迁移和侵袭。使用 RNA 纯化染色质分离、染色质免疫沉淀和双荧光素酶报告基因测定研究了 m6A 修饰的 LEAWBIH 的机制。LEAWBIH 高表达并与 HCC 患者的不良生存相关。 LEAWBIH 被鉴定为 m6A 修饰的转录本。 m6A 修饰增加了 LEAWBIH 转录物的稳定性。 LEAWBIH 的 m6A 修饰水平在 HCC 中增加，LEAWBIH 的高 m6A 修饰水平预示生存率较差。 LEAWBIH 以 m6A 修饰依赖性方式促进 HCC 细胞增殖、迁移和侵袭。机制研究表明，m6A 修饰的 LEAWBIH 激活 Wnt/β-catenin 信号传导。 m6A 修饰的 LEAWBIH 与 m6A 阅读器 YTHDC1 结合，进一步与 H3K9me2 去甲基化酶 KDM3B 相互作用并将 H3K9me2 去甲基化酶 KDM3B 募集至 CTNNB1 启动子，导致 H3K9me2 去甲基化和 CTNNB1 转录激活。功能性救援试验表明，阻断 Wnt/β-catenin 信号传导消除了 LEAWBIH 在 HCC 中的作用。m6A 修饰的 LEAWBIH 通过表观遗传激活 Wnt/β-catenin 信号传导在 HCC 中发挥致癌作用，强调 m6A 修饰的 LEAWBIH 作为 HCC 的一个有前途的治疗靶点HCC.© 2023 Wei 等人。

N6-methyladenosine (m6A) modification plays an important role in regulating RNA maturation, stability, and translation. Thus, m6A modification is involved in various pathophysiological processes including hepatocellular carcinoma (HCC). However, the direct contribution of m6A modifications to RNA function in HCC remains unclear. Here, we identified LEAWBIH (long non-coding RNA epigenetically activating Wnt/β-catenin signalling in HCC) as an m6A-modified long non-coding RNA (lncRNA) and investigated the effects of m6A on the function of LEAWBIH in HCC.Quantitative polymerase chain reaction was performed to measure the gene expression in tissues and cells. The level of m6A modification was detected using a methylated RNA immunoprecipitation assay and single-base elongation- and ligation-based qPCR amplification method. Cell proliferation was evaluated using the Glo cell viability and CCK-8 assays. Cell migration and invasion were evaluated using Transwell migration and invasion assays. The mechanisms of m6A modified LEAWBIH were investigated using chromatin isolation by RNA purification, chromatin immunoprecipitation, and dual-luciferase reporter assays.LEAWBIH was highly expressed and correlated with poor survival in HCC patients. LEAWBIH was identified as a m6A-modified transcript. m6A modification increased LEAWBIH transcript stability. The m6A modification level of LEAWBIH was increased in HCC, and a high m6A modification level of LEAWBIH predicted poor survival. LEAWBIH promotes HCC cell proliferation, migration, and invasion in an m6A modification-dependent manner. Mechanistic investigations revealed that m6A-modified LEAWBIH activated Wnt/β-catenin signaling. m6A-modified LEAWBIH binds to the m6A reader YTHDC1, which further interacts with and recruits H3K9me2 demethylase KDM3B to CTNNB1 promoter, leading to H3K9me2 demethylation and CTNNB1 transcription activation. Functional rescue assays showed that blocking Wnt/β-catenin signaling abolished the role of LEAWBIH in HCC.m6A-modified LEAWBIH exerts oncogenic effects in HCC by epigenetically activating Wnt/β-catenin signaling, highlighting m6A-modified LEAWBIH as a promising therapeutic target for HCC.© 2023 Wei et al.