Background <p>Laryngeal squamous cell carcinoma (LSCC) is the most common type of head and neck malignancy. NAT10 is a catalytic enzyme for ac4C and is involved in the progression of a variety of cancers. This study aimed to explore the effects and potential mechanisms of NAT10 in LSCC.</p> Methods <p>Pyroptosis was assessed by measuring the release of lactic dehydrogenase, pyroptosis rate, and pyroptosis-related proteins. The RNA and protein levels were detected by quantitative real-time PCR and western blot, respectively. Potential mechanisms were validated using flow cytometry, ac4C dot blot, methylated RNA immunoprecipitation (MeRIP), RIP, and Dual-Luciferase Reporter Assay experiments.</p> Results <p>The result showed that the levels of NAT10 in LSCC tissues and cells were elevated and positively correlated with tumor grading and clinical staging. Knockdown of NAT10 promoted the pyroptosis of LSCC cells. NAT10 directly interacted with ELANE, suppressed the stability of the ELANE mRNA. NAT10 inhibited pyroptosis in LSCC by downregulating the ELANE expression in vivo and in vitro.</p> Conclusion <p>NAT10 inhibited the pyroptosis of LSCC cells and contributed to LSCC progression by suppressing ELANE mRNA stability in ac4C modification manner, indicating that the NAT10-ac4C-ELANE axis might be a potential target for LSCC.</p>