NAT10-mediated ac4C acetylation of PKM2 promotes papillary thyroid carcinoma through regulating glycolysis.

Papillary thyroid carcinoma (PTC) represents the most common histological variant among differentiated thyroid cancers. N4-acetylcytidine (ac4C) is a modified nucleoside in mRNA, which is essential for the regulation of mRNA stability. N-acetyltransferase (NAT)10 is an enzyme responsible for ac4C modification, acting as an "ac4C writer." This study aimed to investigate the role of NAT10 in PTC and the underlying mechanisms involved. A total of forty-eight patients with PTC were recruited, and a nude mouse model was developed. Reverse transcription quantitative polymerase chain reaction (RT-qPCR) was employed to assess the expression levels of NAT10 and pyruvate kinase (PK) M2. Cell viability was evaluated using the cell counting kit-8 (CCK-8), while proliferation was determined through colony formation assay. Glycolysis indicators were measured using commercially available kits. Bioinformatics analysis was used to screen the target genes of NAT10. The interaction between NAT10 and PKM2 was analyzed via RNA immunoprecipitation (RIP) and dual luciferase reporter assays. NAT10 was increased in PTC tissues and cells. NAT10 inhibition suppressed cell proliferation and glycolysis in PTC cells and restrained tumor growth and tissue malignancy in xenograft tumor mice. Mechanistically, PKM2 has been identified as a downstream regulatory target influenced by NAT10-mediated ac4C acetylation. Additionally, rescue experiments indicated that the overexpression of PKM2 enhanced both cell viability and glycolysis activity in PTC cells. NAT10-mediated ac4C acetylation of PKM2 exacerbated the progression of PTC by promoting glycolysis, which might provide a new insight for PTC therapy.