Abstract: Objective To investigate the synergistic induction of cytotoxicity and apoptosis by the combination of tumor necrosis factor related apoptosis inducing ligand (TRAIL), adriamycin(ADM) and IFN-γ in osteosarcoma MG-63 cells. Methods MTT assays were used to evaluate the cytotoxic effects of MG-63, and flow cytometric analyses were used for the assessment of apoptosis rate after different treatments. Results The combined use of the three treatments induced significantly higher apoptosis and cytotoxicity than exposure to TRAIL, ADM or IFN-γ alone, but the combination of two of the three agents did not show synergistic effects. Conclusion The combination of TRAIL, ADM and IFN-γ could significantly increase cytotoxic and apoptosis effects on MG-63 cells.