Abstract
Melanoma cancer is the most aggressive skin cancer type with poor prognosis. Chemotherapy have been used in the past in treating melanoma cancer, however, the method usually results in toxicity. Therefore, a call has been made to develop cancer treatments using medicinal plants to reduce drug toxicity. The aim of this study was to determine the efficacy of Cannabis sativa extracts as co-treatment for melanoma cancer. The hypothesis of this study was that Cannabis sativa extracts has potential for usage as a cotreatment for melanoma cancer. Results obtained supported the hypothesis since down regulation of apoptotic genes in melanoma cancer was observed after cotreating with Cannabis sativa extracts. The IC50 values of Cannabis sativa ethanol and methanol extracts were obtained at 51.31315 μg/ml and 57.34135 μg/ml respectively for A375 cells. Normal cells were used to check if there are any serious cytotoxic effects, IC50 values for Cannabis sativa ethanol and methanol extracts were obtained at 79.577850 μg/ml and 59.5754 μg/ml respectively. RMG-1 cells showed IC50 at 84.748 μg/ml and 34.66475 μg/ml for Cannabis sativa ethanol and methanol extracts respectively. CYTOSMART LUX3 bright field microscope was used for morphological analysis of both A375 and RMG-1 cells and changes were observed after treating with Cannabis sativa ethanol and methanol extracts. Apoptosis analysis to determine mode of cell death was done using CYTOSMART fluorescence microscope. Early apoptotic effects were observed in both Cannabis sativa ethanol and methanol extracts on A375 cells. Early apoptotic effects were also observed after treating RMG-1 cells with ethanol extract. Caspase activity was done on both A375, and RMG-1 cells and caspase 3/7 activity were increased after treating with Cannabis sativa ethanol and methanol extracts. RT-PCR showed that RBBP6 and BCl2 was down-regulated after treating with Cannabis sativa ethanol and methanol extracts. SiRNA co-treatment with ethanol and methanol extracts resulted in low expression of both RBBP6 and BCl2. P53 was upregulated after treatment with the extracts and co-treatment with the extracts and siRNA.
Key words: Melanoma cancer, A375, ethanol extract, methanol extract, RBBP6.