Fibroblast growth factor 2 and forskolin induce mineralization-associated genes in two kinds of osteoblast-like cells.

Fibroblast growth factor 2 (FGF2) and cyclic AMP (cAMP) play critical roles in controlling the differentiation of osteoblasts and mineralization of bone. We have previously reported that each of FGF2 and forskolin (FSK) alone increase transcription of the bone sialoprotein (BSP) gene, and that together (FGF/FSK) they upregulate BSP gene expression synergistically in rat osteoblast-like ROS 17/2.8 cells. However, other genes that are upregulated after stimulation by FGF2, FSK or FGF/FSK remain unclear. In the present study, we investigated candidate genes associated with mineralization after stimulation by FGF2, FSK and FGF/FSK in two kinds of osteoblast-like cells using microarray and real-time PCR. In ROS17/2.8 cells, FGF2 and FSK each increased the gene expression of c-FOS (7.2-fold and 10.7-fold, respectively). However, FGF/FSK did not induce c-FOS gene expression. FGF2 increased the expression of the dentin matrix protein 1 (DMP1, 129.8-fold) gene. In contrast, FGF/FSK increased the expression of the amphiregulin (AREG, 73-fold) gene maximally. In human osteoblast-like Saos2 cells, FGF2 increased the expression of the osteopontin (SPP1, 16.7-fold), interleukin-8 (IL8, 6.4-fold) and IL11 (4.8-fold) genes. FSK induced the expression of the IL6 (2.6-fold), IL11 (4.0-fold), chemokine ligand 13 (CXCL13, 2.8-fold) and bone morphogenetic protein 2 (BMP2, 2.5-fold) genes. These results suggest that FGF2 and FSK might be crucial regulators of mineralization and bone formation.