Antiproliferative effect of selected zingiberaceae species against human oral cancer cells / Muhammad Amirul Amil

Muhammad Amirul , Amil (2024) Antiproliferative effect of selected zingiberaceae species against human oral cancer cells / Muhammad Amirul Amil. Masters thesis, Universiti Malaya.

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Abstract

Oral squamous cell carcinoma (OSCC) is a prevalent and highly challenging form of oral cancer that poses significant health burdens worldwide. Current treatment options for OSCC are limited and often associated with adverse side effects. Therefore, there is an urgent need to explore alternative therapeutic approaches. Natural compounds derived from medicinal plants have gained significant attention for their promising prospects as effective and safe antiproliferative agents. This study aims to investigate the in vitro antiproliferative effects of essential oils and extracts obtained from Curcuma aeruginosa, Curcuma mangga, Curcuma xanthorrhiza, Kaempferia galanga, and Zingiber zerumbet on OSCC cells. The essential oil of the rhizomes obtained by hydrodistillation were subjected to chemical analyses using a combination of GC/MS and GC-FID analytical techniques. Substantial percentages of identified compounds were found, such as β- myrcene (79.77%) in the essential oil of C. mangga, followed by zerumbone (48.39%), ethyl-cinnamate (40.14%), and β-curcumene (34.90%) in the essential oils of Z. zerumbet, C. xanthorrhiza, and K. galanga, respectively. The essential oils and extracts of the Zingiberaceae species were evaluated for their antiproliferative effects against OSCC namely H103 and ORL-204 cancer cells; and hGF a noncancerous human gingival fibroblast cells using the MTT assay. Notably, the hexane fractionated extract of C. mangga (IC50 = 10.72 μg/mL), ethyl acetate fractionated extract of C. xanthorrhiza (IC50 = 4.99 μg/mL), and Z. zerumbet’s fractionated extract in hexane and essential oil (IC50 = 1.93 and 8.29 μg/mL, respectively) displayed significant antiproliferative activity on H103 cancer cells. Moreover, the crude methanol extract of C. mangga (IC50 = 5.04 μg/mL), ethyl acetate fractionated extract of C. xanthorrhiza (IC50 = 3.07 μg/mL), and the hexane fractionated extract as well as the essential oil of Z. zerumbet (IC50 = 1.13 and 8.21 μg/mL, respectively), demonstrated potent inhibition against the proliferation of ORL-204 cells. Based on the observed low IC50 value of the essential oil of Z. zerumbet in the MTT viability assay, subsequent investigations were conducted to explore the cell death pathway and the underlying mechanisms of ORL-204 cells treated with the essential oil of Z. zerumbet. These investigations encompassed in vitro morphological examinations, assessment of mitochondrial membrane potential, detection of caspases (caspase-3, -8, and -9), analysis of DNA fragmentation-TUNEL, and cell cycle analysis. Consistently, these assays confirmed the occurrence of apoptosis within the ORL-204 cells upon treatment with the essential oil of Z. zerumbet, suggesting its ability to induce programmed cell death and inhibit cell proliferation. Altogether, the findings of this study highlight the antiproliferative effects of the essential oil of Z. zerumbet on OSCC cells and its ability to induce programmed cell death through both extrinsic and intrinsic pathways. These results suggest that the essential oil of Z. zerumbet has the capacity to become a valuable antiproliferative agent in the future treatment of oral cancer, opening up possibilities for further research and development of novel therapeutic approaches.

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