Shahbaz, Khushboo
(2020)
Cytotoxic effect of phoenix dactylifera L. (ajwa date extract) on oral squamous cell carcinoma cell line.
Masters thesis, Universiti Sains Malaysia.
Abstract
Oral squamous cell carcinoma (OSCC) is a major public health problem
globally. The prognosis for patients with OSCC remains poor despite the advancement
in treatment modalities and diagnostic technology. Therefore, there is an urgent need
to explore natural products that effectively cure this disease. Ajwa date fruit, was
reported to have strong anti-oxidant and anti-cancer properties. However, no study has
focused on Ajwa date flesh and pits extract effect on OSCC yet. The aim of the current
study is to investigate the cytotoxic and apoptotic potential of Ajwa date flesh (ADF)
and Ajwa date pit (ADP) extract on HSC-2 cell line. ADF and ADP were extracted
with solvent extraction method using hexane, acetone and ethanol, which were then
subjected to antioxidant assay by 2,2-diphenyl-1-picrylhydrazyl (DPPH), 3-(4,5-
Dimethylthiazol-2-yl)-2, 5-Diphenyltetrazolium Bromide (MTT) assay for
cytotoxicity assessment. For apoptosis assessment, Annexin-V apoptosis detection kit
for 24, 48 and 72 hours was used. In this study, acetone extracts of ADF and ADP had
the highest radical scavenging and antioxidant activities followed by the ethanolic
extracts. Whereas, ADP appeared to have significantly higher antioxidant effects than
the ADF. In this study, the MTT assay demonstrated that acetone extracts of ADF and
ADP were significantly cytotoxic against HSC-2 cells in a dose and time-dependent
manner. The half inhibitory concentration (IC50) of ADF was found to be 8.69 mg/ml
at 24h and the maximum cell growth inhibition was observed at 50 mg/ml. The IC50
for the ADP was found to be 0.97 mg/ml at 24h and the maximum cell growth
inhibition was observed at 5 mg/ml. Statistical analysis of the flowcytometry assay
showed that the treatment with ADF and ADP extracts had significant apoptotic effect
which occurred in a dose-dependent manner. At treatment with higher doses of ADP
and ADF extract HSC-2 cells were seen in late apoptotic stage. ADP extract
demonstrated higher apoptotic activity than ADF extract. In addition, combined
treatment of ADF and ADP were also performed on HSC-2 cells which demonstrated
higher apoptotic activity when compared to the single extract. In conclusion, Ajwa
date fruit has promising cytotoxic effect by inhibiting growth and proliferation of
OSCC cells and inducing cell death by apoptosis. Furthermore, high radical
scavenging activity of ADP will promote it as a potential candidate for further research
to isolate the antioxidant compounds and to be used in combination with the fruit flesh
in the development of anti-cancer drug in future.
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