- Short Communication
Osteoblast differentiation of NIH3T3 fibroblasts is associated with changes in the IGF-I/IGFBP expression pattern
Cellular & Molecular Biology Letters volume 11, pages 461–474 (2006)
Insulin-like growth factors (IGFs) and IGF-binding proteins (IGFBPs) are essential regulators for osteoblast proliferation and differentiation. It has been reported that Dexamethasone (Dex), an active glucocorticoid (GC) analogue, synergizes the stimulatory effect of 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) on osteoblast differentiation in the mouse fibroblastic cell line NIH3T3. I investigated whether this stimulatory effect is associated with changes in the expression pattern of the IGF/IGFBP system. Quantitative real-time PCR technology was used to quantify the gene expression levels of the IGF-system during osteoblast differentiation and in response to 1,25(OH)2D3 or Dex alone under serum-containing and serum-free culture conditions. Interestingly, NIH3T3 was shown to express high mRNA levels of IGF-I, IGF-II and IGFBP-5, and low levels of both IGFBP-2 and-6. During osteoblast differentiation (days 6-12), IGF-I mRNA was repressed by more than 60%, while the transcript of IGFBP-5 was markedly up-regulated, by more than 50-fold. Similarly, treatment with Dex alone resulted in a dose-and time-dependent increase in the expression of IGFBP-5 and a decrease in IGF-I mRNA. Treatment with 1,25(OH)2D3 alone increased the mRNA levels of IGF-I and IGFBP-6 by around 4-and 7-fold, respectively, in a dose-and time-dependent manner. In conclusion, my data demonstrated that osteoblast differentiation of NIH3T3 is associated with changes in the expression pattern of IGFs/IGFBPs, which are regulated by glucocorticoid in the presence of 1,25(OH)2D3. Modulation of the IGF/IGFBP levels by glucocorticoid might suggest important roles for the IGF-system in mediating the osteoblast differentiation of the NIH3T3 cell line.
Bone morphogenetic protein-2
Core binding factor
- runt domain:
alfa subunit 1
insulin like growth factor binding proteins
insulin like growth factors
mesenchymal stem cells
Kassem, M., Kristiansen, M. and Abdallah, B.M. Mesenchymal stem cells: cell biology and potential use in therapy. Basic Clin. Pharmacol. Toxicol. 95 (2004) 209–214.
Langdahl, B.L., Kassem, M., Moller, M.K. and Eriksen, E.F. The effects of IGF-I and IGF-II on proliferation and differentiation of human osteoblasts and interactions with growth hormone. Eur. J. Clin. Invest. 28 (1998) 176–183.
Rajaram, S., Baylink, D.J. and Mohan, S. Insulin-like growth factor-binding proteins in serum and other biological fluids: regulation and functions. Endocr. Rev. 18 (1997) 801–831.
Mohan, S. and Baylink, D.J. IGF-binding proteins are multifunctional and act via IGF-dependent and-independent mechanisms. J. Endocrinol. 175 (2002) 19–31.
Robson, H., Siebler, T., Shalet, S.M. and Williams, G.R. Interactions between GH, IGF-I, glucocorticoids, and thyroid hormones during skeletal growth. Pediatr. Res. 52 (2002) 137–147.
Eijken, M., Koedam, M., van Driel, M., Buurman, C.J., Pols, H.A. and van Leeuwen, J.P. The essential role of glucocorticoids for proper human osteoblast differentiation and matrix mineralization. Mol. Cell Endocrinol. 248 (2006) 87–93.
Delany, A.M. and Canalis, E. Transcriptional repression of insulin-like growth factor I by glucocorticoids in rat bone cells. Endocrinology 136 (1995) 4776–4781.
Cheng, S.L., Zhang, S.F., Mohan, S., Lecanda, F., Fausto, A., Hunt, A.H., Canalis, E. and Avioli, L.V. Regulation of insulin-like growth factors I and II and their binding proteins in human bone marrow stromal cells by dexamethasone. J. Cell Biochem. 71 (1998) 449–458.
Gurlek, A., Pittelkow, M.R. and Kumar, R. Modulation of growth factor/cytokine synthesis and signaling by 1alpha,25-dihydroxyvitamin D(3): implications in cell growth and differentiation. Endocr. Rev. 23 (2002) 763–786.
Kveiborg, M., Flyvbjerg, A., Eriksen, E.F. and Kassem, M. 1,25-Dihydroxyvitamin D3 stimulates the production of insulin-like growth factor-binding proteins-2,-3 and-4 in human bone marrow stromal cells. Eur. J. Endocrinol. 144 (2001) 549–557.
Nakao, Y., Hilliker, S., Baylink, D.J. and Mohan, S. Studies on the regulation of insulin-like growth factor binding protein 3 secretion in human osteosarcoma cells in vitro. J. Bone Miner. Res. 9 (1994) 865–872.
Shui, C. and Scutt, A.M. Mouse embryo-derived NIH3T3 fibroblasts adopt an osteoblast-like phenotype when treated with 1alpha,25-dihydroxyvitamin D(3) and dexamethasone in vitro. J. Cell Physiol. 193 (2002) 164–172.
Wang, E.A., Israel, D.I., Kelly, S. and Luxenberg, D.P. Bone morphogenetic protein-2 causes commitment and differentiation in C3H10T1/2 and 3T3 cells. Growth Factors 9 (1993) 57–71.
Xiao, Z.S., Hinson, T.K. and Quarles, L.D. Cbfa1 isoform overexpression upregulates osteocalcin gene expression in non-osteoblastic and pre-osteoblastic cells. J. Cell Biochem. 74 (1999) 596–605.
Abdallah, B.M., Haack-Sorensen, M., Fink, T. and Kassem, M. Inhibition of osteoblast differentiation but not adipocyte differentiation of mesenchymal stem cells by sera obtained from aged females. Bone 39 (2006) 181–188.
Koch, H., Jadlowiec, J.A. and Campbell, P.G. Insulin-like growth factor-I induces early osteoblast gene expression in human mesenchymal stem cells. Stem Cells Dev. 14 (2005) 621–631.
Barnes, P.J. Corticosteroids: the drugs to beat. Eur. J. Pharmacol. 533 (2006) 2–14.
Canalis, E., Bilezikian, J.P., Angeli, A. and Giustina, A. Perspectives on glucocorticoid-induced osteoporosis. Bone 34 (2004) 593–598.
Smith, E., Coetzee, G.A. and Frenkel, B. Glucocorticoids inhibit cell cycle progression in differentiating osteoblasts via glycogen synthase kinase-3beta. J. Biol. Chem. 277 (2002) 18191–18197.
Jorgensen, N.R., Henriksen, Z., Sorensen, O.H. and Civitelli, R. Dexamethasone, BMP-2and 1,25-dihydroxyvitamin D enhance a more differentiated osteoblast phenotype: validation of an in vitro model for human bone marrow-derived primary osteoblasts. Steroids 69 (2004) 219–226.
Minuto, F., Palermo, C., Arvigo, M. and Barreca, A.M. The IGF system and bone. J. Endocrinol. Invest. 28 (2005) 8–10.
Chen, T.L., Mallory, J.B. and Hintz, R.L. Dexamethasone and 1,25(OH)2 vitamin D3 modulate the synthesis of insulin-like growth factor-I insteoblast-like cells. Calcif. Tissue Int. 48 (1991) 278–282.
Chevalley, T., Strong, D.D., Mohan, S., Baylink, D. and Linkhart, T.A. Evidence for a role for insulin-like growth factor binding proteins in glucocorticoid inhibition of normal human osteoblast-like cell proliferation. Eur. J. Endocrinol. 134 (1996) 591–601.
Conover, C.A., Clarkson, J.T. and Bale, L.K. Effect of glucocorticoid on insulin-like growth factor (IGF) regulation of IGF-binding protein expression in fibroblasts. Endocrinology 136 (1995) 1403–1410.
Fielder, P.J., Tauber, J.P., Wilson, K.F., Pham, H.M. and Rosenfeld, R.G. Insulin-like growth factors (IGFs) stimulate and dexamethasone inhibits IGF binding protein (BP)-5 expression in a mouse pituitary cell line. Growth Regul. 3 (1993) 226–234.
Firth, S.M. and Baxter, R.C. Cellular actions of the insulin-like growth factor binding proteins. Endocr. Rev. 23 (2002) 824–854.
Miyakoshi, N., Richman, C., Kasukawa, Y., Linkhart, T.A., Baylink, D.J. and Mohan, S. Evidence that IGF-binding protein-5 functions as a growth factor. J. Clin. Invest. 107 (2001) 73–81.
Durant, D., Pereira, R.M. and Canalis, E. Overexpression of insulin-like growth factor binding protein-5 decreases osteoblastic function in vitro. Bone 35 (2004) 1256–1262.
Jia, D. and Heersche, J.N. Expression of insulin-like growth factor system constituents in differentiating rat osteoblastic cell populations. Growth Horm. IGF. Res. 12 (2002) 399–410.
Chen, T.L., Chang, L.Y., Bates, R.L. and Perlman, A.J. Dexamethasone and 1,25-dihydroxyvitamin D3 modulation of insulin-like growth factor-binding proteins in rat osteoblast-like cell cultures. Endocrinology 128 (1991) 73–80.
Chenu, C., Valentin-Opran, A., Chavassieux, P., Saez, S., Meunier, P.J. and Delmas, P.D. Insulin like growth factor I hormonal regulation by growth hormone and by 1,25(OH)2D3 and activity on human osteoblast-like cells in short-term cultures. Bone 11 (1990) 81–86.
Linkhart, T.A. and Keffer, M.J. Differential regulation of insulin-like growth factor-I (IGF-I) and IGF-II release from cultured neonatal mouse calvaria by parathyroid hormone, transforming growth factor-betaand 1,25-dihydroxyvitamin D3. Endocrinology 128 (1991) 1511–1518.
Schmid, C., Schlapfer, I., Gosteli-Peter, M.A., Hauri, C., Froesch, E.R. and Zapf, J. 1 alpha,25-dihydroxyvitamin D3 increases IGF binding protein-5 expression in cultured osteoblasts. FEBS Lett. 392 (1996) 21–24.
Kim, E.J., Schaffer, B.S., Kang, Y.H., Macdonald, R.G. and Park, J.H. Decreased production of insulin-like growth factor-binding protein (IGFBP)-6 by transfection of colon cancer cells with an antisense IGFBP-6 cDNA construct leads to stimulation of cell proliferation. J. Gastroenterol. Hepatol. 17 (2002) 563–570.
Schmid, C. Insulin-like growth factors. Cell Biol. Int. 19 (1995) 445–457.
Yan, T., Wergedal, J., Zhou, Y., Mohan, S., Baylink, D.J. and Strong, D.D. Inhibition of human osteoblast marker gene expression by retinoids is mediated in part by insulin-like growth factor binding protein-6. Growth Horm. IGF. Res. 11 (2001) 368–377.
Bach, L.A. IGFBP-6 five years on; not so ‘forgotten’? Growth Horm. IGF. Res. 15 (2005) 185–192.
About this article
Cite this article
Abdallah, B.M. Osteoblast differentiation of NIH3T3 fibroblasts is associated with changes in the IGF-I/IGFBP expression pattern. Cell Mol Biol Lett 11, 461–474 (2006). https://doi.org/10.2478/s11658-006-0038-y
- Gene expression
- Osteoblast differentiation