Ferroptosis and radiotherapy-related genes in head and neck squamous cell carcinoma: diagnostic and prognostic significance

BACKGROUND: Head and Neck Squamous Cell Carcinoma (HNSCC) is an aggressive malignancy with poor prognosis and low five-year survival rates. Ferroptosis, an iron-dependent form of cell death, has emerged as a potential factor in cancer therapy. This study aimed to identify differentially expressed genes (DEGs) associated with HNSCC, ferroptosis, and radiotherapy, and to develop a diagnostic model to establish novel biomarkers, early diagnosis and individualized therapy. METHODS: We analyzed TCGA-HNSCC, GSE6631, and GSE107591 datasets to identify Ferroptosis- and Radiotherapy-Related Differentially Expressed Genes (F&RRDEGs).Ferroptosis & radiotherapy scores were calculated, followed by immune infiltration and immunotherapy analyses. A Cox prognostic model was evaluated using nomograms, calibration curves, and decision curve analysis. Real-time quantitative PCR (RT-qPCR) and Western blotting (WB) were used for experimental validation. RESULTS: 11 F&RRDEGs were identified classifying two subtypes related to HNSCC.Significant differences were found in immune cell infiltration and immunophenoscore in CTLA4 and PD1/PD-L1/PD-L2 + CTLA4 between high and low ferroptosis & radiotherapy scores(F&Rs) groups. The 1-year calibration curve of the prognostic risk model showed better and more accurate performance and performance for the five-year prediction. RT-qPCR identified significant differences in the genes ALOX12, EMP1, PIM1, and CA9 between the control and HNSCC groups, with WB confirming radiotherapy-induced upregulation of VDAC1/TFRC (pro-ferroptosis) and downregulation of GPX4/SLC7A11/FTH1 (anti-ferroptosis) in irradiated HNSCC cells. CONCLUSION: ALOX12, EMP1, PIM1, and CA9 exhibited promising potential as diagnostic biomarkers. This study applies immune infiltration analysis to HNSCC research, offering fresh perspectives on patient-specific treatment plans, and accelerating the creation of focused therapies.