Evaluation of the antiproliferative effect of Zingiber zerumbet essential oil against oral squamous cell carcinoma

• Zingiber zerumbet oil was rich in zerumbone (48.39 %), humulene (17.05 %), and camphene (5.83 %). • Zingiber zerumbet oil showed strong cytotoxicity against H103 and ORL-204 cells. • Apoptosis in ORL-204 cells was confirmed by mitochondrial membrane potential loss, caspase activation, and TUNEL-positive DNA fragmentation. Background Oral squamous cell carcinoma (OSCC) treatment necessitates the immediate exploration of alternative therapies due to the limitations and side effects of current methods. This study aims to investigate the antiproliferative effects of Zingiber zerumbet essential oil, an ethnobotanically significant species traditionally used in folk medicine, against OSCC. Methods An overview of the biological activities associated with the essential oils and extracts from Zingiber zerumbet was provided to validate their potential for therapeutic and commercial applications. The essential oil was extracted using the hydrodistillation method. GC/MS and GC-FID analyses were employed to identify the chemical components in the essential oil. Investigations involved MTT viability tests, in vitro morphological examinations, assessment of mitochondrial membrane potential, detection of caspase-3, -8, and -9 activities, analysis of DNA fragmentation using TUNEL, and cell cycle assays. Results A total of 18 chemical components in the essential oil were determined using a combination of GC/MS and GC-FID analyses, in which zerumbone (48.39%), humulene (17.05%), and camphene (5.83 %) were identified as major components. The essential oil of Zingiber zerumbet inhibited the growth of OSCC, namely H103 and ORL-204 cancer cells (IC 50 = 1.93 and 1.13 µg/mL, respectively) in the MTT viability assay. The ORL-204 cell line was used for subsequent investigations, exploring the underlying cell death mechanisms and cell cycle phase distribution. All assays consistently confirmed the occurrence of apoptosis and distinct cell cycle phase distributions in ORL-204 cells upon treatment with the essential oil of Zingiber zerumbet . Conclusion Altogether, the essential oil of Zingiber zerumbet demonstrates preliminary antiproliferative activity against oral cancer cells, opening possibilities for further research and development of novel therapeutic approaches.