목적: High recurrence rates in head and neck squamous cell carcinoma (HNSCC) significantly affect prognosis, especially in radioresistant HNSCC (RR-HNSCC). Non-thermal plasma (NTP) therapy can effectively suppress the progression of HNSCC; however, the therapeutic mechanism of NTP therapy for RR-HNSCC remains unclear. In this study, we investigated the regulatory role of NTP in the RR-HNSCC signaling pathway and identified its signature genes. 방법:After constructing two RR-HNSCC cell lines, we prepared cell lysates from cells treated or not treated with non-thermal plasma-activated media (NTPAM) and performed RNA sequencing to determine their mRNA expression profiles. Based on the RNA sequencing results, we identified differentially expressed genes, followed by bioinformatics analysis to identify candidate molecules potentially associated with NTPAM therapy for RR-HNSCC. 결과:NTPAM decreased RR-HNSCC cell viability in vitro. The RNA sequencing results indicated that NTPAM treatment activated the reactive oxygen species pathway and induced ferroptosis in RR- HNSCC cell lines. Among the 1924 genes correlated with radiation treatment, eight showed statistical significance in both the cell lines and The Cancer Genome Atlas (TCGA) cohort. Only five genes, ABCC3, DUSP16, PDGFB, RAF1, and THBS1, showed consistent results between the NTPAM data sequencing and TCGA data. LASSO regression analysis revealed that five genes were associated with cancer prognosis, with a hazard ratio (HR) of 2.26. In RR-HNSCC cells, NTPAM affected DUSP16, PDGFB, and THBS1 as the activated marker within 6 h and persisted for 12 h. Furthermore, enrichment analysis indicated that these three differential genes were associated with ECM, TGF-β, PI3K-AKT, and MET pathways. 결론:NTPAM therapy enhances cytotoxicity in RR-HNSCC cell lines by inducing specific ROS- mediated ferroptosis. DUSP16, PDGFB, and THBS1 were identified as crucial targets for reversing radiation resistance induced by NTPAM therapy, providing insights into the mechanisms and clinical applications of NTPAM treatment in RR-HNSCC.