Abstract: Objective To investigate the expression of Livin in osteosarcoma MG-63 cells and the effect of Livin anti-sense oligonucleotide（ASODN） on its activity and function. Methods The ASODN targeting Livin were designed and constructed. According to the results of transfection efficiency evaluation， the liposome／oligonucleotide complexes were prepared by adding oligonucleotide and then transferred into MG-63 cells. The immunohistochemical staining and Western blotting were used to detect the expression of Livin at 24th h after transfection in MG-63 cells.The proliferation at 72nd h and apoptosis at 24th h after transfection were measured by CellTiter-Glo and Flow cytometry，respectively. Results The Livin was mainly expressed in the cytoplasm of MG-63 cells. We chose ASODN with the highest transfection efficiency to transfer MG-63 cells. The expression of Livin protein decreased significantly after the transfection， and Livin ASODN at different concentrations inhibit the proliferation and migration of MG-63 cells with significant difference. The apoptotic rate of Lip-ASODN group was higher than those of blank control group and negative control group in a dose-dependent manner （P＜0.05）. Conclusion The ASODN targeting Livin can inhibit the proliferative capacity and promote the apoptosis of osteosarcoma MG-63 cells.