Proteomic analysis on effectors involved in BMP-2-induced osteogenic differentiation of beagle bone marrow
Ji-Jie Hu1, Ya-Wei Liu2, Min-Yi He3
1Department of Orthopaedics and Traumatology, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong 510515, China.
Objective:
To identify the protein regulation profile of recombinant human bone morphogenetic protein-2 (rhBMP-2)-induced osteogenic differentiation in beagle bone marrow stem cells (BMSCs).
Methods:
Beagle BMSCs were isolated and cultured with or without rhBMP-2. Two-dimensional gel electrophoresis was used to determine the differences in protein expression in rhBMP-2-induced and non-induced BMSCs. Real-time PCR and western blotting analyses were used to verify the expression patterns of selected proteins.
Results:
After the induction, the osteogenic differentiation of beagle BMSCs was activated successfully. Nine and 11 proteins were found to be down- and up-regulated by rhBMP-2, respectively. The increase in Lim and SH3 domain protein 1(LASP1) and the decrease in ferritin were verified by real-time PCR and western blotting analyses.
Conclusions:
Among the 20 rhBMP-2-regulated factors, there is empirical evidence supporting the involvement of LASP1 and ferritin in osteogenic differentiation. LASP1 plays an important role in the regulation of the activity of the cytoskeleton, and ferritin is an important molecule in cellular iron homeostasis. Further studies focused on these 20 proteins will help elucidate the molecular mechanism(s) through which rhBMP-2 induces osteogenic differentiation of BMSCs.
