Introduction: Cancer is one of the major medical problems threatening human health. Breast cancer is the most prevalent type of cancer in women. For more efficient treatment, new treatment strategies have been evolved based on combination therapy in which two or more different methods are exploited for this aim. In the present study, a combination therapy based on radiotherapy (using neutron radiation emitted from a Am-Be neutron source) and biotherapy (using curcumin) is applied to investigate the treatment efficiency of MCF-7 breast cancer in a three-dimensional culture medium. Materials and methods: MTT, neutral red uptake assay, nitric oxide, glutathione assay, catalase, cytochrome c, comet assay, and caspase-3 were used to determine the effect and type of mortality due to neutron effect as well as the combined effect of neutron and curcumin in cancer cells. Results: The results of cytotoxicity due to neutron irradiation as well as the combined effect of neutron and curcumin showed that neutron irradiation with or without curcumin at 5, 10, 15, and 20 hours reduced the survival of tumor cells. Conclusion: Overall, this study showed that the neutron source Am-Be with the applied doses was able to destroy the co, and ch bonds of curcumin, resulting in cell death and apoptosis induction in breast cancer cells. It was caused by neutron radiation. Due to the significant anti-cancer effect of curcumin in 3D culture, the use of this molecule before or after neutron therapy is recommended.