Integrative multi-omics analysis identifies a core ferroptosis signature and validates resveratrol as a novel inducer in pancreatic cancer.

Pancreatic ductal adenocarcinoma (PDAC) has a dismal prognosis due to treatment resistance and an immunosuppressive, fibrotic microenvironment. Ferroptosis, a novel form of regulated cell death, offers a promising therapeutic strategy. Transcriptomic datasets GSE28735 and GSE62452 were integrated for differential expression and WGCNA. Ferroptosis-related genes were intersected, followed by machine learning (LASSO, SVM-RFE, etc.) to screen core genes. Enrichment, immune infiltration, and single-cell transcriptome (GSE197177) analyses were performed. Clinical validation used HPA and TCGA. Resveratrol effects were assessed by molecular docking, CCK-8, Western blot, and ferroptosis inhibitor rescue assays (GSH measurement, Fer-1 intervention). Twenty-seven PDAC ferroptosis-related genes were identified, and five core genes were refined by machine learning. Single-cell analysis further confirmed six key genes: GPX4, CTSB, NOX4, TFRC, HIF1A, and TGFB1. They were significantly enriched in ferroptosis, HIF-1, and TGF-β pathways. M1/M2 macrophages showed notable infiltration and correlated with gene expression. Clinical validation revealed high CTSB/TFRC protein expression in PDAC; high TGFB1 predicted poor prognosis. In vitro, resveratrol downregulated multiple key proteins, suppressed tumor cell activity, and enhanced ferroptosis sensitivity. Ferroptosis inhibitor (Fer-1) partially reversed resveratrol-induced GSH depletion and protein changes, confirming ferroptosis involvement. This study unveils a ferroptosis regulatory network in PDAC involving HIF-1/TGF-β signaling and macrophage crosstalk. Resveratrol targets this network to induce ferroptosis, offering novel biomarkers and a potential therapeutic strategy for PDAC.