Abstract
IntroductionPhoenix dactylifera L (Date palm) is a native plant of the Kingdom of Saudi Arabia (KSA) and other Middle Eastern countries. Ajwa date has been described in the traditional and alternative medicine to provide several health benefits including anticholesteremic, antioxidant, hepatoprotective and anticancer effects, but most remains to be scientifically validated. Herein, we evaluated the anticancer effects of the Methanolic Extract of Ajwa Date (MEAD) on human breast adenocarcinoma (MCF7) cells in vitro.MethodsMCF7 cells were treated with various concentrations (5, 10, 15, 20 and 25 mg/ml) of MEAD for 24, 48 and 72 h and changes in cell morphology, cell cycle, apoptosis related protein and gene expression were studied.ResultsPhase contrast microscopy showed various morphological changes such as cell shrinkage, vacuolation, blebbing and fragmentation. MTT (2-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay demonstrated statistically significant dose-dependent inhibitions of MCF7 cell proliferation from 35% to 95%. Annexin V-FITC and TUNEL assays showed positive staining for apoptosis of MCF7 cells treated with MEAD (15 mg and 25 mg for 48 h). Flow cytometric analyses of MCF7 cells with MEAD (15 mg/ml and 20 mg/ml) for 24 h demonstrated cell cycle arrest at 'S' phase; increased p53, Bax protein expression; caspase 3activation and decreased the mitochondrial membrane potential (MMP). Quantitative real time PCR (qRT-PCR) analysis showed up-regulation of p53, Bax, Fas, and FasL and down-regulation of Bcl-2.ConclusionsMEAD inhibited MCF7 cells in vitro by the inducing cell cycle arrest and apoptosis. Our results indicate the anticancer effects of Ajwa dates, which therefore may be used as an adjunct therapy with conventional chemotherapeutics to achieve a synergistic effect against breast cancer.
Highlights
Phoenix dactylifera L (Date palm) is a native plant of the Kingdom of Saudi Arabia (KSA) and other Middle Eastern countries
Annexin V-FITC and TUNEL assays showed positive staining for apoptosis of MCF7 cells treated with Methanolic Extract of Ajwa Date (MEAD) (15 mg and 25 mg for 48 h)
Our results indicate the anticancer effects of Ajwa dates, which may be used as an adjunct therapy with conventional chemotherapeutics to achieve a synergistic effect against breast cancer
Summary
MCF7 cells were treated with various concentrations (5, 10, 15, 20 and 25 mg/ml) of MEAD for 24, 48 and 72 h and changes in cell morphology, cell cycle, apoptosis related protein and gene expression were studied. Ajwa dates that were fresh, ripe, medium sized, fleshy, soft, and have basal white lines on the black exocarp was used in the present study. Originally cultivated in the city of AlMadina Al-Munawwarah were purchased from “Tamar Market” of Al-Madina Al-Munawwarah, KSA. MEAD was prepared using previously described protocol with slight modification [17]. The edible part of date fruit was manually separated, freeze dried and coarsely powdered using pestle and mortar. The powdered contents were extracted in methanol (1:3 ratio, weight to volume) on a shaking incubator (Human Lab, Gyeonggi-Do, Korea) at 25°C for 48 h. The supernatant was separated and freeze dried using a lyophilizer (ilShin Biobase Europe, The Netherlands). The freeze dried powder was aliquoted and stored at -80°C until use in the experiments
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