Caffeine is a commonly consumed psychoactive substance that has been shown to have various effects on cellular processes, including cell growth and survival. In this study, we investigated the potential of Caffeine to enhance the cytotoxic effects of two commonly used chemotherapeutic agents, doxorubicin and oxaliplatin, on B16F10 cells. We evaluated the cytotoxicity, calculated the IC50 and combination index of the medications, estimated the cell cycle of the cells, and evaluated the apoptotic and anti-apoptotic genes through gene expression analysis. Our results demonstrated that Caffeine significantly potentiated the cytotoxicity of both doxorubicin and oxaliplatin, resulting in a more significant reduction in cell viability compared to treatment with the chemotherapeutic agents alone. Additionally, Caffeine also enhanced the pro-apoptotic effects of the chemotherapeutic agents, leading to increased levels of apoptosis in the B16F10 cells. The combination of doxorubicin and oxaliplatin with Caffeine was found to be synergistic. These findings suggest that caffeine may have the potential to improve the efficacy of current chemotherapeutic regimens and may offer a novel approach to enhance cancer treatment.