NAT10-Mediated N4-Acetylation of SLC7A11 mRNA Promotes Hepatocellular Carcinoma Progression and Metastasis by Repressing Ferroptosis

Abstract

N-acetyltransferase 10 (NAT10) catalyzes N4-acetylcytidine (ac4C) modification of mRNA, yet its role in hepatocellular carcinoma (HCC) and ferroptosis remains elusive. A retrospective study with 100 HCC patients assessed NAT10 and SLC7A11 expression. Immunohistochemistry evaluated protein levels in HCC tissues. In vitro experiments using HCC cell lines explored NAT10's impact on proliferation, migration, invasion, and apoptosis. In vivo, nude mice with HCC xenografts underwent stable si-NAT10 transfection, monitoring tumor growth and metastasis. Ferrostatin-1 treatment investigated NAT10's role in ferroptosis. RNA immunoprecipitation and acRIP assays examined NAT10's influence on SLC7A11 mRNA stability. Elevated NAT10 expression correlated with poor outcomes and TACE therapy resistance in HCC. NAT10 silencing inhibited HCC cell functions, promoting apoptosis and ferroptosis in vitro; Ferrostatin-1 reversed these effects. In vivo models confirmed NAT10's significance in HCC progression, reducing tumor volume and weight with increased apoptosis. Ferrostatin-1 counteracted these effects, underlining ferroptosis in NAT10-driven tumorigenesis. NAT10 knockdown increased Fe²⁺ accumulation and decreased SLC7A11 and GPX4 expression, indicating that NAT10 knockdown enhanced ferroptosis. Mechanistically, NAT10 stabilized SLC7A11 mRNA via ac4C acetylation, suppressing ferroptosis. This epigenetic modification emerged as a regulator of ferroptosis evasion in HCC. Overall, NAT10 promotes HCC progression by inhibiting ferroptosis through ac4C acetylation of SLC7A11 mRNA.