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Objective: To study the inhibition of expression of Survivin gene by synthesized short Survivin-siRNA in osteosarcoma cell line MG-63. Methods: Chemically synthesized short Surviving-siRNA was transfected into osteosarcoma cell line MG-63. The Survivin mRNA and protein level were detected by reverse transcription-polymerase chain reaction (RT-PCR) and flow cytometry (FCM). The biological morphology and growth inhibition of MG-63 were observed with light microscopy and MTT assay, respectively. Results: Short siRNA targeting Survivin down-regulated the transcription and the protein level of Survivin oncogene. The proliferation of osteosarcoma cell line MG-63 was inhibited after transfection. Conclusion: Chemically synthesized short Survivin-siRNA can effectively inhibit Survivin expression and cell proliferation inosteosarcoma cell line MG-63 Survivin-siRNA mediated Survivin gene silencing may be a useful therapeutic strategy for osteosarcoma.