Background: Cancer is a serious issue that has a significant effect on the health of all human communities. Tongue cancer is one of the most common head and neck cancers in the world. In the medical world, cancer therapy is a major challenge. Nowadays, natural compounds are important resources of many anti-cancer medications. Venom from honey bees possesses potent anti-cancer effects. Cisplatin is a chemotherapeutic drug that has been used for decades to treat cancer cells. Recently, Combination therapy has been a popular treatment choice for cancer patients.This study aimed to investigate the synergistic cytotoxic effect of honey bee venom (BV) and cisplatin on tongue squamous cell carcinoma cell line (SCC-25).Methods: The cytotoxic effect was determined using Methyl Thiazol Tetrazolium (MTT) assay, microscopic examination, P53 and caspase-3 were quantified by Real-Time Polymerase chain reaction (RT-PCR) and statistical analysis.Results: The findings revealed that the tested drugs' cytotoxic potential against SCC-25 cells is dose dependent. The half-maximal inhibitory concentration (IC50) value of MTT assay of BV/cisplatin mix decreased significantly compared to IC50 values of BV and cisplatin in sole formulation. Microscopic examination showed that BV and cisplatin alone and in combination mainly produced apoptotic cell death. Regarding RT-PCR results, P53 and caspase-3 expression were significantly increased in SCC-25-treated cells (P= 0.0001).Conclusions: The combined use of BV and cisplatin induced marked synergistic cytotoxic effect on SCC-25 cell line.