The present study aimed to investigate the effect of Dermatopontin (DPT) gene silencing on the apoptosis and proliferation of osteosarcoma MG‑63 cells. Three eukaryotic expression vectors of short hairpin (sh)RNA fragments targeting different loci of DPT were designed and transfected into an osteosarcoma cell line MG‑63. The cells were assigned to a blank, shRNA‑control, DPT‑shRNA‑a, DPT‑shRNA‑b or DPT‑shRNA‑c group. The shRNA with the highest silencing efficiency was screened using reverse transcription‑quantitative polymerase chain reaction and western blotting. The screened shRNA was transfected into MG‑63 cells. The proliferation, cell cycle and apoptosis of MG‑63 cells were measured using a Cell Counting Kit‑8 assay, flow cytometry and Annexin V‑fluorescein isothiocyanate assay. The recombinant plasmids containing DPT shRNA were successfully constructed. DPT gene silencing was able to significantly reduce the proliferation rate of MG‑63 cells (P<0.05). The proportion of cells in the G0/G1 phase and in the G2/M phase increased significantly (both P<0.05), while the proportion of cells in the S phase decreased (P<0.05). Furthermore, the cell apoptosis rate increased significantly (P<0.05). These results demonstrate that DPT gene silencing is able to reduce the proliferation of MG‑63 cells, slow down cell cycle progression and promote apoptosis, hence may become a novel target for the treatment of osteosarcoma.