Stretch-induced PTH-related protein gene expression in osteoblasts

Abstract

Mechanical forces play a critical role in regulating skeletal mass and structure. We report that mechanical loading induces PTHrP in osteoblast-like cells and that TREK-2 stretch-activated potassium channels seem to be involved in this induction. Our data suggest PTHrP as a candidate endogenous mediator of the anabolic effects of mechanical force on bone. Introduction: Mechanical force has anabolic effects on bone. The PTH-related protein (PTHrP) gene is known to be mechanically inducible in smooth muscle cells throughout the organism, and N-terminal PTH and PTHrP products have been reported to have anabolic effects in bone. We explored the idea that PTHrP might be a candidate mediator of the effects of mechanical force on bone. Materials and Methods: Mechanical loading was applied by swelling osteoblast-like cells in hypotonic solution and/or by application of cyclical stretch through a FlexerCell apparatus. RNase protection assay and real-time quantitative PCR analysis were used to assay PTHrP gene expression. Results and Conclusion: Stretching UMR201-10B osteoblast-like cells by swelling in hypotonic solutions rapidly increased PTHrP mRNA. This induction was insensitive to gadolinium and nifedipine, to the removal of extracellular calcium, and to depletion of endoplasmic reticulum calcium, indicating that neither stretch-activated cation channels, L-type calcium channels, nor ER calcium is involved in the induction of PTHrP. The TREK family potassium channels are activated by both stretch and intracellular acidosis, and we identified these channels in osteoblast-like cells by PCR. Intracellular acidification increased PTHrP mRNA expression in UMR-201-10B cells, and siRNA targeted against the TREK-2 gene reduced endogenous TREK-2 expression and dampened PTHrP mRNA induction. Cyclical stretch also induced PTHrP in UMR-201-10B osteoblast-like cells and in MLO-A5 post-osteoblast-pre-osteocyte cells, the latter a stage in the osteoblastic differentiation program that is likely to be a key target of force in vivo. Our evidence suggests PTHrP as a candidate mediator of the anabolic effects of mechanical force on bone. © 2005 American Society for Bone and Mineral Research.