Chen Ming, Yang Yuan, Hu Guangsheng, Peng Zhong, Wen Wu
Department of Gastroenterology and Hepatology, the First Affiliated Hospital, Hengyang Medical School, University of South China, Hengyang City, 421001, Hunan, China.
Department of Gastroenterology and Hepatology, Yiyang Central Hospital, Yiyang City, 413099, Hunan, China.
Ann Hepatol. 2025 Jan 3;30(2):101776. doi: 10.1016/j.aohep.2025.101776.
Deregulation of mA methylation, the most prevailing RNA modification, participates in cancer pathogenesis. METTL16, an atypical methyltransferase, functions as a pro-tumorigenic factor in hepatocellular carcinoma (HCC). Here, we explored the action of METTL16 on HCC glycolysis and the associated mechanism.
Expression analysis was done by quantitative PCR, immunoblotting, or immunohistochemistry. Cell sphere formation, invasiveness, apoptosis, proliferation and viability were detected by sphere formation, transwell, flow cytometry, EdU and CCK-8 assays, respectively. Xenograft studies were performed to analyze the role in vivo. Methylated RNA immunoprecipitation (MeRIP) and RIP assays were used to verify the METTL16/PFKM relationship. PFKM mRNA stability was tested by actinomycin D treatment. Chromatin immunoprecipitation (ChIP) and luciferase assays were performed to analyze the POU3F2/METTL16 relationship.
In HCC, METTL16 expression was elevated, and increased levels of METTL16 transcript predicted poor HCC prognosis. METTL16 deficiency resulted in suppressed HCC cell growth, invasiveness and sphere formation. Moreover, METTL16 depletion diminished HCC cell glycolysis. Mechanistically, PFKM expression was positively associated with METTL16 expression. METTL16 mediated m6A methylation to stabilize PFKM mRNA via an IGF2BP3-dependent manner. Restored PFKM expression exerted a counteracting effect on METTL16 deficiency-mediated in vitro cell phenotype alterations and in vivo xenograft growth suppression. Furthermore, POU3F2 promoted the transcription of METTL16 in HCC cells.
Our findings define the crucial role of the POU3F2/METTL16/PFKM axis in HCC pathogenesis, offering the potential opportunity to combat HCC.
m⁶A甲基化作为最普遍的RNA修饰，其失调参与癌症发病机制。METTL16是一种非典型甲基转移酶，在肝细胞癌（HCC）中作为促肿瘤因子发挥作用。在此，我们探究了METTL16对HCC糖酵解的作用及相关机制。
通过定量PCR、免疫印迹或免疫组化进行表达分析。分别采用成球实验、Transwell实验、流式细胞术、EdU实验和CCK-8实验检测细胞成球能力、侵袭能力、凋亡、增殖和活力。进行异种移植研究以分析其在体内的作用。采用甲基化RNA免疫沉淀（MeRIP）和RIP实验验证METTL16/PFKM关系。用放线菌素D处理检测PFKM mRNA稳定性。进行染色质免疫沉淀（ChIP）和荧光素酶实验分析POU3F2/METTL16关系。
在HCC中，METTL16表达升高，METTL16转录水平升高预示着HCC预后不良。METTL16缺陷导致HCC细胞生长、侵袭和成球能力受到抑制。此外，METTL16缺失减少了HCC细胞糖酵解。机制上，PFKM表达与METTL16表达呈正相关。METTL16介导m⁶A甲基化以依赖IGF2BP3的方式稳定PFKM mRNA。恢复PFKM表达对METTL16缺陷介导的体外细胞表型改变和体内异种移植生长抑制具有拮抗作用。此外，POU3F2促进HCC细胞中METTL16的转录。
我们的研究结果确定了POU3F2/METTL16/PFKM轴在HCC发病机制中的关键作用，为对抗HCC提供了潜在机会。
