Diosgenin Enhances the Sensitivity to Methotrexate Through Oxidative DNA Damage in Saos-2 Osteosarcoma Cancer Cells

Previously, diosgenin, a steroidal saponin, has demonstrated therapeutic potential for osteosarcoma. However, the underlying mechanisms still remain unknown.This study was designed to investigate the impact of diosgenin on methotrexate-mediated apoptosis in Saos-2 cells, specifically related to DNA damage.To assess the cell vitality of Saos-2 cells, we treated them with methotrexate, diosgenin, and a mixture of both, then performed the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide technique. Western blot was used to evaluate DNA damage by measuring the expression of γ-H2AX protein. The ELISA method was also applied to find the quantity of 8-oxo-2'-deoxyguanosine, and flow cytometry was used to analyze apoptosis.Combining methotrexate with diosgenin led to a major reduction in cell proliferation rate compared to monotreatments (<i>p</i><0.05). This combination significantly increased apoptosis, measured by flow cytometry, and increased levels of γ-H2AX protein and 8-oxo-2'-deoxyguanosine.Diosgenin significantly augmented methotrexate-mediated apoptosis in Saos-2 cells through enhanced DNA damage mechanisms. These findings suggest that diosgenin could serve as a promising adjuvant therapeutic agent to improve the efficacy of current methotrexate-based chemotherapy regimens in osteosarcoma treatment, potentially reducing required drug doses and minimizing associated toxicity while maintaining therapeutic effectiveness.