Abstract:

BACKGROUND: Brg1 is an ATPase subunit of the SWI/SNF complex and plays an important role in the regulation of gene transcription, replication and recombination, skeletal muscle differentiation and inhibition carcinogenesis.
OBJECTIVE: To explore the Brg1 regulation mechanism in bone morphogenetic protein 2 (BMP-2)-induced osteogenic differentiation.
METHODS: Mouse calvarial osteoblasts were primarily cultured through collagenase digestion and induced differentiation using 0, 50 or 200 μg/L BMP2 to select the optimal dose. Transcription time dynamics analysis of Brg1 was enforced by RT-PCR and Western blot after BMP-2 treatments; ALP staining was used to evaluate the capability of osteogenic differentiation after BMP-2 treatments. The role of BMP-2 on Brg1 mRNA expression level was detected by quantitative RT-PCR and the protein level was detected by Western Blot. Effect of Brg1 on BMP-2-induced differentiation was analyzed by Brg1-siRNA; Ad-Dlx5 was obtained by recombinant adenovirus.
RESULTS AND CONCLUSION: BMP-2 could induce the osteogenic differentiation of primarily cultured mouse osteoblast, and the optimal BMP2 induction condition was 200 μg/L. The expression of Brg1 was up-regulated by BMP-2 in primarily cultured mouse osteoblasts. BMP-2 induced the osteogenic differentiation was inhibited by Brg1-siRNA. The expression of Dlx5 was regulated by Brg1. Brg1 could regulate the BMP-2-induced osteogenic differentiation of primarily cultured mouse osteoblast through regulating the expression of Dlx5.