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The purpose of the present study was to evaluate the effects of resveratrol and shikonin, two natural agents, on the induction and maintenance of osteosarcoma cell apoptosis. The results showed that resveratrol and shikonin were able to induce apoptosis in osteosarcoma cell lines in a dose-dependent manner, and induced a more potent cytotoxic effect in the MG63 cell line. DAPI staining revealed that resveratrol induced a slight nuclear fragmentation in MG63 cells, while the treatment of MG63 cells with shikonin induced typical apoptotic nuclear changes, such as chromatin condensation and increased fragmentation. Following treatment, the MG63 cells exhibited an increase in the expression of Bax, Fas, caspase-3 and p53, but an decrease in Bcl-2 expression. In addition, the activities of caspase-3 were significantly enhanced after incubation with resveratrol and shikonin. 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