Abstract
Introduction Melanoma cancer is the most aggressive skin cancer type with a poor prognosis. Chemotherapy has 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 establish the role of RBBP6 in melanoma cancer development and progression. The hypothesis of this study was that RBBBP6 is highly expressed in melanoma cancer and serves as an early biomarker. Results obtained supported the hypothesis since high expression of RBBP6 in melanoma cancer was observed in untreated control melanoma cells. Methodology 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT), Cytosmart fluorescence, and quantitative Polymerase Chain Reaction (qPCR) were used. Results The IC 50 values of Cannabis sativa EtOH and MeOH extracts were obtained at 51.31315 and 57.34135 µg/mL, respectively, for A375 cells. Normal cells were used to check if there are any serious cytotoxic effects, IC 50 values for Cannabis sativa EtOH and MeOH extracts were obtained at 79.577850 and 59.5754 µg/mL, respectively. RMG-1 cells showed IC 50 at 84.748 and 34.66475 µg/mL for Cannabis sativa EtOH and MeOH 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 EtOH and MeOH extracts. Apoptosis analysis to determine the mode of cell death was done using a CYTOSMART fluorescence microscope. Early apoptotic effects were observed in both Cannabis sativa EtOH and MeOH extracts on A375 cells. Early apoptotic effects were also observed after treating RMG-1 cells with EtOH extract. Caspase 3/7 activity was done on both A375 and RMG-1 cells, and caspase 3/7 activity were increased after treating with Cannabis sativa EtOH and MeOH extracts. Real Time Polymerase Chain Reaction (RT-PCR) showed that RBBP6 and BCl 2 were down-regulated after treating with Cannabis sativa EtOH and MeOH extracts. Conclusion siRNA co-treatment with EtOH and MeOH extracts resulted in low expression of both RBBP6 and BCl 2 . P53 was upregulated after treatment with the extracts and co-treatment with the extracts and siRNA.