N6-甲基腺嘌呤（m6A）修饰在调控mRNA命运中起着重要作用。非编码RNA在包括肝细胞癌（HCC）在内的多种疾病中也具有重要作用。然而，m6A修饰对非编码RNA的潜在影响仍不清楚。在本研究中，我们鉴定了一种新的m6A修饰的ATP8B1-AS1，并旨在研究m6A对ATP8B1-AS1在HCC中的表达和作用的影响。我们运用qPCR技术测量相关基因的表达。使用公共数据库分析基因表达与预后之间的关系。利用MeRIP和单碱基延伸联接型qPCR扩增方法测定m6A修饰水平。使用体外和体内功能测定方法研究ATP8B1-AS1在HCC中的作用。通过ChIRP和ChIP实验探究ATP8B1-AS1作用机制。ATP8B1-AS1在HCC组织和细胞系中高表达。ATP8B1-AS1的高表达与HCC患者总体生存差相关。ATP8B1-AS1发生m6A修饰，且已确定ATP8B1-AS1的792位点为m6A修饰位点。m6A修饰增加了ATP8B1-AS1转录本的稳定性。ATP8B1-AS1的m6A修饰水平在HCC组织和细胞系中增加，且与HCC患者总体生存差相关。ATP8B1-AS1促进HCC细胞增殖、迁移和侵袭，而通过突变m6A修饰的792位点可以抑制这些作用。机制研究揭示了m6A修饰的ATP8B1-AS1与m6A阅读子YTHDC1和组蛋白去甲基化酶KDM3B相互作用并招募到MYC启动子区域，在MYC启动子区域降低H3K9me2水平并激活MYC转录。功能救援实验表明，MYC的去除在很大程度上抑制了ATP8B1-AS1的致癌作用。ATP8B1-AS1的m6A修饰水平在HCC中增加并与不良预后相关。通过表观遗传学机制激活MYC表达，m6A修饰的ATP8B1-AS1在HCC中发挥致癌作用。© 2023 Tan et al.

N6-methyladenosine (m6A) modification has shown critical roles in regulating mRNA fate. Non-coding RNAs also have important roles in various diseases, including hepatocellular carcinoma (HCC). However, the potential influences of m6A modification on non-coding RNAs are still unclear. In this study, we identified a novel m6A-modified ATP8B1-AS1 and aimed to investigate the effects of m6A on the expression and role of ATP8B1-AS1 in HCC.qPCR was performed to measure the expression of related genes. The correlation between gene expression and prognosis was analyzed using public database. m6A modification level was measured using MeRIP and single-base elongation- and ligation-based qPCR amplification method. The roles of ATP8B1-AS1 in HCC were investigated using in vitro and in vivo functional assays. The mechanisms underlying the roles of ATP8B1-AS1 were investigated by ChIRP and ChIP assays.ATP8B1-AS1 is highly expressed in HCC tissues and cell lines. High expression of ATP8B1-AS1 is correlated with poor overall survival of HCC patients. ATP8B1-AS1 is m6A modified and the 792 site of ATP8B1-AS1 is identified as an m6A modification site. m6A modification increases the stability of ATP8B1-AS1 transcript. m6A modification level of ATP8B1-AS1 is increased in HCC tissues and cell lines, and correlated with poor overall survival of HCC patients. ATP8B1-AS1 promotes HCC cell proliferation, migration, and invasion, which were abolished by the mutation of m6A-modified 792 site. Mechanistic investigation revealed that m6A-modified ATP8B1-AS1 interacts with and recruits m6A reader YTHDC1 and histone demethylase KDM3B to MYC promoter region, leading to the reduction of H3K9me2 level at MYC promoter region and activation of MYC transcription. Functional rescue assays showed that depletion of MYC largely abolished the oncogenic roles of ATP8B1-AS1.m6A modification level of ATP8B1-AS1 is increased and correlated with poor prognosis in HCC. m6A-modified ATP8B1-AS1 exerts oncogenic roles in HCC via epigenetically activating MYC expression.© 2023 Tan et al.