SP1-mediated NAT10 upregulation drives oral squamous cell carcinoma progression via ac4C-dependent SLC1A5...

Oral squamous cell carcinoma (OSCC) is a prevalent malignancy in the head and neck region, accounting for 90% of all oral cancer cases. Despite advances in surgery and chemoradiotherapy, the molecular mechanisms driving OSCC progression remain incompletely understood, limiting the development of effective targeted therapies. Emerging evidence suggests that transcription factor Specificity protein 1 (SP1) and acetyltransferase N-acetyltransferase 10 (NAT10) may contribute to cancer progression, but their role in OSCC and potential regulation of the amino acid transporter solute carrier family 1 member 5 (SLC1A5) remain unclear. This study aimed to systematically investigate the SP1-NAT10-SLC1A5 regulatory network in OSCC development. The mRNA and protein expression levels were determined by reverse transcription-quantitative PCR (RT-qPCR) and western blotting, respectively. Cell proliferation was assessed using MTT and EdU assays, apoptosis was analyzed by flow cytometry, and cell invasion was evaluated using transwell assay. Enzymatic activity kits were employed to quantify key parameters of glutamine (Gln) metabolism. RNA immunoprecipitation (RIP) assay was used to detect the binding of NAT10 and SLC1A5 mRNA. Dual-luciferase reporter assay was performed to validate the direct regulatory interaction between SP1 and the NAT10 promoter region, followed by chromatin immunoprecipitation (ChIP) assay to assess SP1 occupancy on the NAT10 promoter. A mouse xenograft model was established for in vivo analysis. NAT10 expression was elevated in OSCC tissues and cells. NAT10 inhibition suppressed OSCC cell proliferation, invasion, and Gln metabolism and promoted apoptosis. Additionally, NAT10 mediated the ac4C modification of SLC1A5. Overexpression of SLC1A5 attenuated the effects of NAT10 suppression in OSCC cells. SP1 was identified as the upstream activator of NAT10 expression through transcriptional regulation analysis. Moreover, NAT10 inhibition impeded OSCC growth in vivo by regulating SLC1A5. SP1-induced NAT10 upregulation promoted OSCC advancement through ac4C-mediated SLC1A5 modification, providing new insights into OSCC treatment.

The online version contains supplementary material available at 10.1007/s10616-026-01010-x.