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Clinical and Experimental Otorhinolaryngology > Accepted Articles
doi: https://doi.org/10.21053/ceo.2024.00238    [Accepted]
Altered gene expression profiling by non-thermal plasma-activated media treatment in radioresistant head and neck squamous cell carcinoma
Sicong Zheng1 , Yudan Piao2,3 , Seung-Nam Jung3 , Chan Oh1 , Mi Ae Lim3 , QuocKhanh Nguyen1 , Shan Shen1, Se-Hee Park1, Shengzhe Cui1, Shuyu Piao3, Young Il Kim4 , Ji Won Kim5, Ho-Ryun Won1,5 , Jae Won Chang1,3 , Yujuan Shan6 , Lihua Liu6 , Bon Seok Koo1,3
1Department of Medical Science, Chungnam National University College of Medicine, Daejeon, Republic of Korea
2Dental Department, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, 3 East Qingchun Road, Hangzhou, Zhejiang, China
3Department of Otolaryngology-Head and Neck Surgery, Chungnam National University College of Medicine, Daejeon, Republic of Korea
4Department of Radiation Oncology, Chungnam National University Sejong Hospital, Sejong, Republic of Korea
5Department of Otorhinolaryngology-Head and Neck Surgery, Chungnam National University Sejong Hospital, Sejong, Republic of Korea
6Department of Nutrition, Public Health and Management College, Wenzhou Medical University, Wenzhou, Zhejiang, China
Correspondence  Bon Seok Koo ,Tel: +82-42-280-7695, Fax: +82-42-253-4059, Email: bskoo515@cnuh.co.kr
Received: August 14, 2024; Revised: December 16, 2024   Accepted: January 5, 2025.  Published online: January 6, 2025.
ABSTRACT
Objective
High recurrence rates in head and neck squamous cell carcinoma (HNSCC) significantly affect prognosis, especially in radioresistant HNSCC (RR-HNSCC). Nonthermal 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.
Methods
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.
Results
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.
Conclusion
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.
Keywords: Squamous Cell Carcinoma of Head and Neck; Radiotherapy; Plasma Gases; Gene Expression Profiling
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