Abstract
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.
Highlights
Colon cancer is the second most prevalent cause of cancer death in men and women after lung cancer [1]
Mitosox red dye (Invitrogen, Paisley, UK) assay was used to detect the superoxide levels in HCT-116 cell line treated with Z1 and Z2
Our results revealed that the incubation of HCT-116 cells with both Zamzam water (ZW) treatments resulted in morphological changes in HCT-116 cells and inhibited the growth and proliferation of cancer cells
Summary
Colon cancer is the second most prevalent cause of cancer death in men and women after lung cancer [1]. Despite the evolution of scientific methods in understanding the biology of cancer and the discovery of new treatments, the mortality rate is still high. The reason that may explain the low rate of response to treatment is the resistance of cancer cells to radiotherapy and chemotherapy. Standard chemotherapeutic agents are used as a supportive or as a main treatment of cancer. Serious side effects have been reported for some anti-tumor chemotherapeutics. Many studies are testing alternative and natural products to increase the response of the patients to the chemotherapeutic drugs or to overcome the toxic side effects of these drugs. Scientists aim to develop oxygen-reduced water with high pH and mineral combination. A novel example is the electrolyzed-reduced water (ERW) which is widely used for its health benefits. It has been shown to possess anticancer properties such as inhibiting cell proliferation in lung A549 cancer cells [4], inhibiting tumor growth of B16 melanoma in tumor injected mice [5] and enhancing the mitochondrial apoptosis pathway in human promyelocytic leukemia cells (HL-60) [3]
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