Effects of siRNA mediated Cdc2 silencing on MG63 cell proliferation and apoptosis

Abstract

The present study aimed to determine the effect of small interfering RNA (siRNA)-induced inhibition of cyclin-dependent kinase 2 (Cdc2) expression on osteosarcoma MG63 cell proliferation and apoptosis. An siRNA expression plasmid, psilencer 2.1-U6/Cdc2, targeting the Cdc2 gene, and a control psilencer 2.1-U6/Scramble plasmid were constructed and transfected into MG63 cells using liposomes. Cdc2 expression in the MG63 cells was investigated by western blot analysis and real-time polymerase chain reaction. Cell morphology was also examined. The effects of psilencer 2.1-U6/Cdc2 on MG63 cell proliferation and the cell cycle were detected via MTT and flow cytometry, respectively. Expression levels of apoptosis related molecules, B-cell lymphoma 2 (Bcl-2) and Bcl-2-associated X (Bax) were determined by western blot analysis. MG63 cells stably transfected with the psilencer 2.1-U6/Cdc2 plasmid (MG63-siRNA/Cdc2) and negative control cells, MG63-siRNA/Scramble, were successfully obtained. The silencing efficiencies of the Cdc2-expressing mRNA and protein in MG63-siRNA/Cdc2 were 86 and 89% of that of the control MG63-siRNA/Scramble cells, respectively. Interference of Cdc2 expression inhibited MG63 cell proliferation and was demonstrated to significantly increase and decrease cells in the G2/M and S phases, respectively. Cdc2 expression silencing had negligible effects on Bcl-2 and Bax expression in MG63 cells. In conclusion, silencing of Cdc2 expression suppresses proliferation of osteosarcoma MG63 cells but has negligible effects on apoptosis. Copyright © 2013 Spandidos Publications Ltd.