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- Title
Effects of Dlx2 overexpression on osteogenic differentiation of MC3T3-E1 cells.
- Authors
SUN Hao; WANG Xu-dong; SHEN Guo-fang; JIANG Xin-quan; ZHANG Xiu-li; DAI Jie-wen; LU Jing-ting
- Abstract
PURPOSE: To investigate the effect of Dlx2 overexpression on osteogenic differentiation of MC3T3-E1 cells in vitro. METHODS: Dlx2-expression retrovirus vector was constructed by subcloning and verified by sequencing. MC3T3-E1 cells were transfected with pMSCV-Dlx2 and selected with puromycin for stable clones. The expression of Dlx2 was determined by RT-PCR and Western blot. The level of osteogenetic biomarkers ALP, OCN, Runx2 and Msx2 was quantified by real-time PCR. ALP detection and Alizarin red staining were conducted to evaluate the effect of Dlx2 overexpression on osteogenic differentiation. The data were analyzed for ANOVA with SNK method using SAS 6.04 software package. RESULTS: pMSCV-Dlx2 was successfully constructed and verified by direct sequencing and Dlx2 overexpression in vitro. Enhanced ALP activity and Alizarin red staining were observed in MC3T3-E1-Dlx2 cells as compared with the control. During osteogenetic induction, Dlx2 overexpression up-regulated ALP and Msx2 in the early stage, while enhanced OCN expression in the late stage. Runx2 expression was different but not reached statistical significance. CONCLUSION: Dlx2 overexpression induces osteogenic differentiation of MC3T3-E1 cells via up - regulating bone formation-related genes. Supported by Shanghai Leading Academic Discipline Project (S30206), Research Fund of Science and Technology Commission of Shanghai Municipality (10JC1408700), Research Fund of Shenkang Company (SHDC12010205), Research Fund of Shanghai Municipal Health Bureau (2009077) and Combined Engineering and Medicine Project of Shanghai Jiao Tong University (YG2010MS55).
- Subjects
OSTEOSARCOMA; PUROMYCIN; MOLECULAR cloning; REVERSE transcriptase polymerase chain reaction; NUCLEOTIDE sequence; ANALYSIS of variance; BIOMARKERS; RETROVIRUSES
- Publication
China Journal of Oral & Maxillofacial Surgery, 2011, Vol 9, Issue 3, p189
- ISSN
1672-3244
- Publication type
Article