BACKGROUND: Abnormal expression of N-acetyltransferase 10 (NAT10) has been shown to promote the progression of various tumors, including non-small cell lung cancer (NSCLC). This study was designed to investigate the role of NAT10 in NSCLC and the underlying mechanism. METHODS: Reverse transcription-quantitative polymerase chain reaction and Western blot were used to analyze the levels of NAT10 in NSCLC cell lines. The cell viability, proliferation, and apoptosis of A549 and PC9 cell lines were detected by cell counting kit-8, colony formation, and flow cytometry. N4-acetylcytidine (ac RESULTS: NAT10 was increased in NSCLC cell lines. The ac CONCLUSIONS: NAT10 regulated glycolysis and apoptosis in NSCLC via ac