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

Que,Xiang‑Yong; Li,Yi; Han,Yu; Li,Xin‑Zhi

doi:10.3892/mmr.2012.1209

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

Authors:
- Xiang‑Yong Que
- Yi Li
- Yu Han
- Xin‑Zhi Li
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Affiliations: Department of Orthopedics, Renhe Hospital of Three Gorges University, Yichang 443001, P.R. China, Institute of Molecular Biology, Medical College, Three Gorges University, Yichang 443002, P.R. China
Published online on: November 28, 2012 https://doi.org/10.3892/mmr.2012.1209
Pages: 466-470

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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.

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