FOXM1 transcriptionally activates NEIL3 to inhibit ferroptosis in lung adenocarcinoma cells.

Lung adenocarcinoma (LUAD) is responsible for a substantial portion of cancer-related deaths, carrying a bleak treatment outlook. The application of ferroptosis-focused treatments has shown great potential. This research is committed to uncovering the molecular mechanisms by which Nei Like DNA Glycosylase 3 (NEIL3) impacts ferroptosis in LUAD, in the quest for robust biomarkers. Using The Cancer Genome Atlas database, qRT-PCR, and Western blot (WB), we evaluated the expression of NEIL3 in LUAD tissues and cells, then performed gene set enrichment analysis to identify enriched gene sets. Predictive tools hTFtarget and MoLoTool assisted in identifying potential upstream transcription factors and their promoter binding sites for NEIL3, following which we conducted Pearson correlation analysis. The binding affinity of NEIL3 to Forkhead box protein M1 (FOXM1) was validated with dual-luciferase and chromatin immunoprecipitation assays. Cell viability was determined by measuring MDA and Fe(2+) content in cells with the aid of cell counting kit-8. Lipid reactive oxygen species (ROS) levels were detected by flow cytometry, and WB was employed to evaluate the expression of GPX4 and SLC7A11 proteins. An upregulation of NEIL3 is observed in LUAD tissues and cell lines, particularly within pathways linked to ferroptosis. When NEIL3 was knocked down, there was a decline in the viability of LUAD cells, coupled with elevated MDA, Fe(2+), and lipid ROS levels. Protein expression of GPX4 and SLC7A11 was inhibited, but these phenotypes were rescued by the application of a ferroptosis inhibitor. FOXM1 could interact with the NEIL3 gene promoter, initiating its transcription. In the context of LUAD, the activation of NEIL3 by FOXM1 constitutes the FOXM1/NEIL3 axis that counteracts ferroptosis in LUAD cells.