Zamzam water (ZW) is a naturally hard alkaline type of water with unique physical and chemical properties that are different from any other water. The aim of the current work is to evaluate the potential anticancer activity of ZW against colon cancer. Human colon cancer HCT-116 and human skin fibroblast HSF cell lines were treated with two treatment conditions of ZW, Z1 with adjusted pH to 7.4 and Z2 without pH adjustment (pH 8). Cell viability was assessed using MTT and trypan blue dye exclusion assays. Cell cycle alterations and the type of cell death were investigated using flow cytometry technique. Cellular and mitochondrial reactive oxygen species (ROS) levels were quantified by H2DCFDA and MitoSOX assays, respectively. The results of the current study showed that both ZW treatments reduced cell viability of cancer cells. MTT assay showed a significant reduction in cell viability to 87% and 66%, respectively, after treatment with Z1 and Z2 (p≤0.0001). Cell death has occurred via apoptotic pathway under the two treatment conditions. The percentage of early apoptosis was 3%, 3.5% and 2.8% in control, Z1 and Z2 respectively. In the late apoptotic stage, there was a significant increase (4.2%) only for Z2 treatment in comparison to the control (p≤ 0.001). The cells were arrested in the G2/M phase of the cell cycle, and decreased in G1 phase after 24 hours of treatment with ZW. Only Z2 treatment, showed an increase in the production of both cytoplasmic and mitochondrial ROS. In conclusion, our results indicate, for the first time, that ZW induces apoptosis of human colon cancer HCT-116 cells, suggesting the potential anticancer effect of ZW against colon cancer.
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