Assessing the anti-cancer activities of novel therapeutics on human Medulloblastoma cancer cells

Abstract

Background: Medulloblastoma (MB) is the most common malignant intra-cranial solid tumor among children. Extensive research has been implicated in this field to look for novel therapeutic options that could help alleviate patients’ symptoms or at least cease progression of the disease. Of late, new studies have indicated proof that a subpopulation of cells inside the tumor, namely cancer stem cells, is believed to be answerable for the protection from most therapeutics and radiation treatment, resulting in malignancy. Henceforth, it is critical to distinguish and to explicitly target those cancer stem cells. Therefore, assessing the effects of different novel therapeutics may contribute to the advancement of knowledge of the discrete mechanisms leading to pathological changes in the brain function and that can block the growth and spread of cancer by interfering with specific molecules. The overall aim of this proposal is to study the anti-neoplastic effect of thymoquinone which is abundantly present in seeds of Nigella sativa L. that is popularly known as black cumin or black seed, on human medulloblastoma cell lines (D556 and D283). Methods: D556 and D283 will be cultured and maintained in DMEM Ham’s F-12 media with 10% FBS and 1% P/S in a humidified incubator (37°C; 5% CO2). The cells will be treated with different concentrations of Thymoquinone reconstituted in methanol. MTT and Trypan blue exclusion assays will be used to assess the anti-proliferative effect of Thymoquinone in vitro. In addition, the ability of Thymoquinone to inhibit cell migration will be tested using the wound-healing migration assay. Finally, the 3D sphere-formation assay will be used to investigate the effect of Thymoquinone on the cancer stem/progenitor cells population in both cell lines. Preliminary and Anticipated Results: using MTT and trypan blue exclusion assays, our preliminary data suggest that Thymoquinone reduced the proliferation of D556 and D283 cells in a time- and dose-dependent manners. We anticipate that Thymoquinone will inhibit cell migration by inhibiting wound closure compared to non-treated conditions. Furthermore, we expect that Thymoquinone will reduce sphere-formation ability of cells in a dose-dependent manner. Conclusion: this will be one of the first studies to assess the effect of Thymoquinone on human medulloblastoma cells in vitro and based on our preliminary results we believe that the drug might have potential therapeutic value.