A novel positive-feedback loop between CEBPA and NAT10 promotes non-small cell lung cancer progression

Non-small cell lung cancer (NSCLC) severely impairs patients' health and quality of life, placing a substantial burden on both patients' families and society. N4-acetylcytidine (ac4C), a pivotal RNA modification among the most abundant in eukaryotes, regulates disease development. Research has shown the ac4C-related gene NAT10 promotes lung cancer progression, yet the mechanism is unclear. NAT10 expression was assessed using data from the TCGA, CPTAC, ENCORI, and TNMplot databases, as well as qRT-PCR, and Western blot analyses. Cell proliferation was evaluated using MTT and EdU assays. Apoptosis was detected using flow cytomety. Cell invasion and migration were respectively detected by transwell and wound-healing assays. Cancer stem cell properties were analyzed via tumor sphere formation assays. Moreover, dual-luciferase reporter assay, ChIP assay, and RIP assay were performed to analyze the interaction between CEBPA and NAT10. Additionally, mRNA stability was evaluated using actinomycin D assay. The animal model was established to analyze the effects of NAT10 and CEBPA on tumor growth in vivo. NAT10 showed high expression levels in NSCLC tissues and cells. Silencing NAT10 inhibited cell viability, proliferation, invasion, migration, and stemness, while promoting apoptosis in NSCLC cells. The transcription factor CEBPA could bind to NAT10, thereby promoting NAT10 expression. Notably, NAT10 mediated the ac4C modification of CEBPA mRNA, which enhanced CEBPA mRNA stability and accelerated the malignant progression of NSCLC. Finally, NAT10 upregulated CEBPA expression, which in turn promoted tumor growth in vivo. NAT10 catalyzed ac4C acetylation on CEBPA mRNA, enhancing its stability and increasing CEBPA protein expression, thereby promoting the malignant progression of NSCLC. The identification of this feedback loop provides a preclinical rationale for developing NSCLC therapeutic strategies targeting NAT10 or CEBPA.