Regulatory effects of LASP1 and ferritin on rhBMP2-induced osteogenic differentiation of bone marrow mesenchymal stem cells / 中华创伤骨科杂志

ABSTRACT

Objective To clarify the function of LIM and SH3 domain protein-1 (LASP1) and ferritin in rhBMP2-induced osteogenic differentiation of beagle bone marrow mesenchymal stem cells (BMSCs).Methods After BMSCs from 3-18-month-old C57BL/6J mice were cultured adherently for 24 hours,they were subjected to osteogenic differentiation for 7,14 and 21 days in 3 groups.BMP2 (100 μg/L) and osteogenic differentiation medium was added in the experimental group,only osteogenic differentiation medium was added in the control group,and nothing was added in the blank group.Osteoblast differentiation was determined by examining marker genes (Runx2,OSX,OCN and OPN) using qRT-PCR.The protein expression of both LASP1 and ferritin was investigated using western blotting.After LASP1 and ferritin were silenced in the cells in the experimental group after transfection of shRNA to target LASP1(m),rhBMP2-induced osteogenesis was repeated to verify the roles of LASP1 and ferritin in osteoblast differentiation.Results The qRT-PCR showed successful osteoblast differentiation in the experimental group.Western blotting verified significant down-regulation of LASP1 and up-regulation of ferritin in the experimental group.After the LASP1 gene was silenced,the expression levels of osteoblast differentiation marker genes in the experimental group were higher than those in the control group.Conclusions rhBMP2 can induce mouse BMSCs to differentiate into osteoblasts in a significant manner.Combined with our preliminary research,the present study may confirm that LASP1 and ferritin,which play an important role in regulating cytoskeleton activity and iron metabolism,are critical in the osteogenic differentiation of mouse BMSCs induced by rhBMP2.