biostudies-literatureJin XNatural Science Foundation of Guangdong Province (Guangdong Natural Science Foundation)National Natural Science Foundation of China (National Science Foundation of China)466-479https://www.ebi.ac.uk/biostudies/studies/S-EPMC11894169biostudies-literatureUnknown32(3)Oxaliplatin-based therapeutics is a widely used treatment approach for hepatocellular carcinoma (HCC) patients; however, drug resistance poses a significant clinical challenge. Epigenetic modifications have been implicated in the development of drug resistance. In our study, employing siRNA library screening, we identified that silencing the m⁶A writer METTL3 significantly enhanced the sensitivity to oxaliplatin in both in vivo and in vitro HCC models. Further investigations through combined RNA-seq and non-targeted metabolomics analysis revealed that silencing METTL3 impeded the pentose phosphate pathway (PPP), leading to a reduction in NADPH and nucleotide precursors. This disruption induced DNA damage, decreased DNA synthesis, and ultimately resulted in cell cycle arrest. MecCell death and differentiationMETTL3 confers oxaliplatin resistance through the activation of G6PD-enhanced pentose phosphate pathway in hepatocellular carcinoma.PMC118941692021A151501070782202905821726468217301681972227Duan JJin XChen JCai MLv YWang FBi JLu LXie DDai MMa CLi SBie FXu SZhou JfalseMETTL3 confers oxaliplatin resistance through the activation of G6PD-enhanced pentose phosphate pathway in hepatocellular carcinoma.Oxaliplatin-based therapeutics is a widely used treatment approach for hepatocellular carcinoma (HCC) patients; however, drug resistance poses a significant clinical challenge. Epigenetic modifications have been implicated in the development of drug resistance. In our study, employing siRNA library screening, we identified that silencing the m⁶A writer METTL3 significantly enhanced the sensitivity to oxaliplatin in both in vivo and in vitro HCC models. Further investigations through combined RNA-seq and non-targeted metabolomics analysis revealed that silencing METTL3 impeded the pentose phosphate pathway (PPP), leading to a reduction in NADPH and nucleotide precursors. This disruption induced DNA damage, decreased DNA synthesis, and ultimately resulted in cell cycle arrest. Mec2025-01-01T00:00:00Z2025 Mar2025-04-22T20:51:26.135Z2025-04-06T03:20:24.69ZS-EPMC118941693947269210.1038/s41418-024-01406-2