Abstract

BACKGROUND Non-small cell lung cancer (NSCLC) progression is tightly linked to tumor microenvironment (TME) remodeling. Taraxerol, a natural compound, has shown potential antitumor activity, but its molecular mechanisms in regulating NSCLC, particularly in TME-related progression, remain poorly characterized. METHODS Single-cell RNA sequencing (scRNA-seq) was conducted to characterize cellular heterogeneity. Network pharmacology integrated differential gene analysis and taraxerol target prediction to identify key regulatory nodes. Molecular interactions were verified via molecular docking and molecular dynamics (MD) simulations. CCK-8, EdU, flow cytometry, and Transwell were employed to assess cell proliferation, apoptosis, migration, and invasion. Xenograft models were established to elucidate taraxerol's function in vivo. Subsequently, tumor-derived exosomes (TDEs) were isolated and characterized by transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA), and Western blot. Macrophage characteristics were validated via qRT-PCR. The interaction between hypoxia-inducible factor 1 alpha subunit (HIF1A) and NIMA-related kinase 6 (NEK6) was investigated via chromatin immunoprecipitation (ChIP) and dual-luciferase reporter assays. RESULTS scRNA-seq revealed NSCLC-specific cellular remodeling, with distinct heterogeneity in macrophage subsets and altered proportions of tumor epithelial cells. HIF1A was a core target of taraxerol, confirmed by molecular docking (binding affinity -8.5 kcal/mol) and MD simulations showing stable interactions. Taraxerol inhibited HIF1A expression in A549/H1299 cells, suppressing proliferation, migration, invasion, and in vivo NSCLC tumorigenesis while promoting apoptosis. Taraxerol-treated TDEs reduced M2 polarization and migration of macrophages, while HIF1A overexpression restored these effects.