Induction of RANKL gene expression by cyclic mechanical strain in osteoblastic MC3T3-E1 cells

  • SAKAI Toshio
    Section of Orthodontics, Department of Oral Growth & Development, Division of Clinical Dentistry, Fukuoka Dental College

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  • 骨芽細胞様細胞MC3T3-E1細胞におけるRANKL遺伝子の周期的細胞伸縮刺激による発現誘導

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The receptor activator of nuclear factor-κB ligand (RANKL), macrophage colony stimulating factor (M-CSF) and osteoprotegerin (OPG) produced by osteoblasts are the important regulatory molecules in osteoclast formation and function. This study examined the effect of cyclic mechanical strain on mRNA levels of RANKL, M-CSF and OPG, and the induction mechanism of RANKL mRNA by the cyclic mechanical strain in osteoblastic MC3T3-E1 cells using RT-PCR. MC3T3-E1 cells plated in α-MEM with 10% FBS on type I collagen coated Bioflex plates were applied to the cyclic mechanical strain at 6 cycles/minute through a Flexercell strain unit. An application of 9% strain force for 2 days increased significantly the RANKL mRNA expression in MC3T3-E1 cells. The cyclic mechanical strain (0-18%) increased the RANKL mRNA expression in a force-dependent manner. In contrast, M-CSF and OPG mRNAs were not affected. The cyclic mechanical strain simultaneously induced the expression of cyclo-oxygenase 2 (COX-2) mRNA and PGE_2 production in MC3T3-E1 cells. Indomethacin, an inhibitor of COX, prevented the increasing of PGE_2 production, but if did not completely inhibit the up-regulated RANKL mRNA expression through The cyclic mechanical strain. These findings suggest that osteoblasts respond to a cyclic mechanical strain and increased the expression of RANKL mRNA, which induction is in part mediated via PGE_2 produced by COX-2.

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