FOXA1‐NAT10 Positive Feedback Pathway Promotes Non‐Small Cell Lung Cancer Cell Progression

Non-small cell lung cancer (NSCLC) is one of the most frequent cancer types and is responsible for the majority of cancer-related deaths all over the world. N-acetyltransferase 10 (NAT10) has been reported to regulate N4-acetylcytidine (ac4C) modification of downstream mRNAs through certain pathways, thereby promoting the progression of diverse tumors, including lung cancer. However, the role and potential mechanism of NAT10 on NSCLC development is still unclear. In this study, GEPIA and TNMplot databases were applied to analyze Forkhead box A1 (FOXA1) expression in lung cancer patients. FOXA1 and NAT10 expression were determined using real-time quantitative polymerase chain reaction (RT-qPCR) and western blot. Cell viability, proliferation, apoptosis, invasion, migration, and stemness were detected using 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2-H-tetrazolium bromide (MTT), 5-ethynyl-2'-deoxyuridine (EdU), flow cytometry, Transwell, wound healing, and sphere formation assays. The abundance of ac4C on FOXA1 mRNA was measured using acRIP-qPCR. Their interaction was verified using RIP and dual-luciferase reporter assays. After JASPAR analysis, the binding between FOXA1 and NAT10 promoter was predicted and then verified using Chromatin immunoprecipitation (ChIP) and dual-luciferase reporter assays. FOXA1 expression was increased in NSCLC tissues and cells, and its knockdown repressed NSCLC cell viability, proliferation, migration, invasion, stemness, and induced apoptosis in vitro. In mechanism, NAT10 maintained FOXA1 mRNA stability in an ac4C-dependent manner. Moreover, FOXA1 was also a transcription factor of NAT10 and increased the transcriptional activity of NAT10 via binding to its promoter region. These findings created a positive feedback cycle to facilitate NSCLC progression. This study elucidated the pivotal role of abnormal activation of the FOXA1/NAT10 positive feedback loop in the malignant progression of NSCLC, thereby uncovering novel prognostic factors and therapeutic targets in NSCLC.