Effects of Dermatopontin gene silencing on apoptosis and proliferation of osteosarcoma MG-63 cells

Abstract

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.