To clarify the role of cementoblast in orthodontic-related root resorption, this study was attempted to examine whether murine cementoblast-like cells are responsive to mechanical stress, and how mechanical forces regulate bone sialoprotein (BSP) and osteopontin (OPN) gene expression in these cells in vitro. In this force-loading model, defined and reproducible mechanical loadings of different magnitudes and types were applied up to 24 h. Besides a transitory and reversible change in cell proliferation, remarkable alterations in gene transcription of BSP and OPN were found. BSP mRNA was suppressed by the stresses. Three and six hours-loadings at 2,000 microstrain up-regulated the expression of OPN mRNA, while the other loadings inhibited it. The study also concluded that 4,000 microstrain was likely to exert more influence on cementoblast-like cells than 2,000 microstrain. Furthermore, no obvious evidence indicated the difference between tension and compression. These results suggested that cementoblast-like cells are sensitive to mechanical stress, and may play a role in regulating orthodontic-related root resorption/repair.