The Src family non-receptor tyrosine kinases (SFK) participate in many aspects of cell function, such as the differentiation, the cell polarity and the migration. Recent studies with gene-targeted mice have revealed that Src plays a key role in osteoclast differentiation, however, the functions of SFK in osteoblasts are not entirely clear at present. In the present studies, we evaluated SFK for regulating the osteoblast functions including cell migration and growth using osteoblast-like MC3T3-E1 cells. Cell migration was significantly blocked in the presence of PP2 (a synthetic Src family kinase specific inhibitor) or LY294002 (a selective inhibitor for PI3-kinase). Further studies demonstrated that PP2 abolished the phosphorylation of AKT and ERK1/2, suggesting the significant role for SFK/AKT/ERK1/2 signaling cascade in the migration of osteoblasts. It is, however, neither PP2 nor LY294002 affected osteoblasts cell proliferation, implicating the signaling cascade involving SFK and PI3-kinase would be distinct from that regulates cell growth. These results suggest that SFK plays a key role in facilitating osteoblasts cell migration by activating AKT and ERK1/2 as potential down-stream targets.