Printed in U.S.A. Copyright © 2000 by The Endocrine Society Expression of Vascular Endothelial Growth Factors and Their Receptors during Osteoblast Differentiation

Endochondral bone formation is regulated by systemically and locally acting growth factors. A role for vascular endothelial growth factor (VEGF) in this process has recently been proposed, because inactivation of VEGF inhibits endochondral bone formation via inhibition of angiogenesis. Despite the known effect of VEGF as specific endothelial growth factor, its effects on osteoblast differentiation have not been studied. We, therefore, examined the expression of VEGF-A, -B, -C, and -D and their receptors in a model of osteoblast differentiation using the mouse preosteoblast-like cell line KS483. Early in differentiation, KS483 cells express low levels VEGF-A, -B, and -D messenger RNA, whereas during mineralization, KS483 cells express high levels. In addition, expression of the VEGF receptors, VEGFR1, VEGFR2, and VEGF165R/neuropilin, coincided with expression of their ligands, being maximally expressed during mineralization. VEGF-A production during osteoblast differentiation was stimulated by insulin-like...

[1]  M. Shibuya,et al.  Increase of vascular endothelial growth factor mRNA expression by 1,25‐dihydroxyvitamin D3 in human osteoblast‐like cells , 1996, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[2]  Mark E. Bolander,et al.  Regulation of Fracture Repair by Growth Factors , 1992, Proceedings of the Society for Experimental Biology and Medicine. Society for Experimental Biology and Medicine.

[3]  R. Nicosia.  What is the role of vascular endothelial growth factor-related molecules in tumor angiogenesis? , 1998, The American journal of pathology.

[4]  H. Fleisch,et al.  Endothelial cells in culture synthesize a potent bone cell active mitogen. , 1986, Endocrinology.

[5]  N. Udagawa,et al.  Cloning of an osteoblastic cell line involved in the formation of osteoclast‐like cells , 1990, Journal of cellular physiology.

[6]  G. Breier,et al.  Expression of vascular endothelial growth factor during embryonic angiogenesis and endothelial cell differentiation. , 1992, Development.

[7]  T. Kitsukawa,et al.  Overexpression of a membrane protein, neuropilin, in chimeric mice causes anomalies in the cardiovascular system, nervous system and limbs. , 1995, Development.

[8]  D. Connolly,et al.  Vascular permeability factor, an endothelial cell mitogen related to PDGF. , 1989, Science.

[9]  B. Olffson.  Vascular endothelial grouth facter B, a novel growth facter for endothelial cells , 1996 .

[10]  Shay Soker,et al.  Neuropilin-1 Is Expressed by Endothelial and Tumor Cells as an Isoform-Specific Receptor for Vascular Endothelial Growth Factor , 1998, Cell.

[11]  G. Rodan,et al.  Expression and regulation of vascular endothelial growth factor in osteoblasts. , 1995, Clinical orthopaedics and related research.

[12]  H. Dvorak,et al.  Tumor cells secrete a vascular permeability factor that promotes accumulation of ascites fluid. , 1983, Science.

[13]  D. Gospodarowicz,et al.  Isolation and characterization of a vascular endothelial cell mitogen produced by pituitary-derived folliculo stellate cells. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[14]  D. Dimitrov,et al.  Identification of the KDR tyrosine kinase as a receptor for vascular endothelial cell growth factor. , 1992, Biochemical and biophysical research communications.

[15]  M. Serio,et al.  Characterization, regulation, and function of specific cell membrane receptors for insulin‐like growth factor I on bone endothelial cells , 1994, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[16]  M. Nimni,et al.  Promotion of calvarial cell osteogenesis by endothelial cells , 1990, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[17]  T. Katagiri,et al.  Subcloning of three osteoblastic cell lines with distinct differentiation phenotypes from the mouse osteoblastic cell line KS-4. , 1996, Bone.

[18]  G. Neufeld,et al.  Vascular endothelial growth factor (VEGF) and its receptors , 1999, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[19]  K. Alitalo,et al.  Vascular endothelial growth factor B, a novel growth factor for endothelial cells. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[20]  D. Maglione.  Persico MG.Isolation of a human placenta cDNA coding for a potein related to the vascular premeability factor , 1991 .

[21]  M. Shibuya,et al.  Neuropilin-1 Is a Placenta Growth Factor-2 Receptor* , 1998, The Journal of Biological Chemistry.

[22]  B. Spiegelman,et al.  Vascular endothelial growth factor. Regulation by cell differentiation and activated second messenger pathways. , 1992, The Journal of biological chemistry.

[23]  P. Chomczyński,et al.  Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. , 1987, Analytical biochemistry.

[24]  R. Ziegler,et al.  Endothelin-1 Is a Potent Regulator of Human Bone Cell Metabolism In Vitro , 1997, Calcified Tissue International.

[25]  M. Shibuya,et al.  The 230 kDa mature form of KDR/Flk-1 (VEGF receptor-2) activates the PLC-γ pathway and partially induces mitotic signals in NIH3T3 fibroblasts , 1997, Oncogene.

[26]  C. Löwik,et al.  IL‐1α, IL‐1β, IL‐6, and TNF‐α Steady‐State mRNA Levels Analyzed by Reverse Transcription‐Competitive PCR in Bone Marrow of Gonadectomized Mice , 1998 .

[27]  S. Oliviero,et al.  Identification of a c-fos-induced gene that is related to the platelet-derived growth factor/vascular endothelial growth factor family. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[28]  俊則 石津谷.  Parathyroid hormone exerts disparate effects on osteoblast differentiation depending on exposure time in rat osteoblastic cells , 1999 .

[29]  A. Uchida,et al.  Suppression of pulmonary metastasis by angiogenesis inhibitor TNP‐470 in murine osteosarcoma , 1995, International journal of cancer.

[30]  K. Alitalo,et al.  A novel vascular endothelial growth factor, VEGF‐C, is a ligand for the Flt4 (VEGFR‐3) and KDR (VEGFR‐2) receptor tyrosine kinases. , 1996, The EMBO journal.

[31]  G S Stein,et al.  Molecular mechanisms mediating proliferation/differentiation interrelationships during progressive development of the osteoblast phenotype. , 1993, Endocrine reviews.

[32]  K. Alitalo,et al.  VEGF-C receptor binding and pattern of expression with VEGFR-3 suggests a role in lymphatic vascular development. , 1996, Development.

[33]  Napoleone Ferrara,et al.  VEGF couples hypertrophic cartilage remodeling, ossification and angiogenesis during endochondral bone formation , 1999, Nature Medicine.

[34]  R. Eddy,et al.  Identification of a new endothelial cell growth factor receptor tyrosine kinase. , 1991, Oncogene.

[35]  J. Seidman,et al.  Structure of wild-type and mutant mouse β 2-microglobulin genes , 1982, Cell.

[36]  M. Kaplan,et al.  THE BIOLOGY OF SAA: IDENTIFICATION OF THE INDUCER, IN VITRO SYNTHESIS, AND HETEROGENEITY DEMONSTRATED WITH MONOCLONAL ANTIBODIES * , 1982, Annals of the New York Academy of Sciences.

[37]  J. Rossant,et al.  Role of the Flt-1 receptor tyrosine kinase in regulating the assembly of vascular endothelium , 1995, Nature.

[38]  N. Ferrara,et al.  The biology of vascular endothelial growth factor. , 1997, Endocrine reviews.

[39]  D. Rifkin,et al.  Autocrine activities of basic fibroblast growth factor: regulation of endothelial cell movement, plasminogen activator synthesis, and DNA synthesis , 1988, The Journal of cell biology.

[40]  P. McSheehy,et al.  Osteoblastic cells mediate osteoclastic responsiveness to parathyroid hormone. , 1986, Endocrinology.

[41]  K. Alitalo,et al.  Localization of VEGF‐B in the mouse embryo suggests a paracrine role of the growth factor in the developing vasculature , 1999, Developmental dynamics : an official publication of the American Association of Anatomists.

[42]  G. Neufeld,et al.  VEGF145, a Secreted Vascular Endothelial Growth Factor Isoform That Binds to Extracellular Matrix* , 1997, The Journal of Biological Chemistry.

[43]  S. Oliviero,et al.  血小板由来増殖因子/血管内皮増殖因子ファミリーと相関するc‐fos誘導性遺伝子の同定 , 1996 .

[44]  Shay Soker,et al.  Characterization of Novel Vascular Endothelial Growth Factor (VEGF) Receptors on Tumor Cells That Bind VEGF via Its Exon 7-encoded Domain (*) , 1996, The Journal of Biological Chemistry.

[45]  J. Winer,et al.  The vascular endothelial growth factor family: identification of a fourth molecular species and characterization of alternative splicing of RNA. , 1991, Molecular endocrinology.

[46]  G. Viglietto,et al.  Isolation of a human placenta cDNA coding for a protein related to the vascular permeability factor. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[47]  C. Patterson,et al.  Enhanced expression of vascular endothelial growth factor in human SaOS-2 osteoblast-like cells and murine osteoblasts induced by insulin-like growth factor I. , 1996, Endocrinology.

[48]  Kanji Sato,et al.  Anabolic effects of 1,25-dihydroxyvitamin D3 on osteoblasts are enhanced by vascular endothelial growth factor produced by osteoblasts and by growth factors produced by endothelial cells. , 1997, Endocrinology.

[49]  S. Nishikawa,et al.  Vascular Endothelial Growth Factor Can Substitute for Macrophage Colony-Stimulating Factor in the Support of Osteoclastic Bone Resorption , 1999, The Journal of experimental medicine.

[50]  Gabriele Bergers,et al.  MMP-9/Gelatinase B Is a Key Regulator of Growth Plate Angiogenesis and Apoptosis of Hypertrophic Chondrocytes , 1998, Cell.

[51]  V. Midy,et al.  Vasculotropin/vascular endothelial growth factor induces differentiation in cultured osteoblasts. , 1994, Biochemical and biophysical research communications.

[52]  D. Merberg,et al.  Molecular cloning of murine FLT and FLT4. , 1993, Oncogene.

[53]  Salvatore Oliviero,et al.  Embryonic expression pattern of the murine figf gene, a growth factor belonging to platelet-derived growth factor/vascular endothelial growth factor family , 1998, Mechanisms of Development.

[54]  H. Phillips,et al.  Developmental expression of binding sites and messenger ribonucleic acid for vascular endothelial growth factor suggests a role for this protein in vasculogenesis and angiogenesis. , 1993, Endocrinology.

[55]  G. Finkenzeller,et al.  1,25-Dihydroxyvitamin D3 induces the expression of vascular endothelial growth factor in osteoblastic cells. , 1997, Endocrine research.

[56]  A. Ullrich,et al.  High affinity VEGF binding and developmental expression suggest Flk-1 as a major regulator of vasculogenesis and angiogenesis , 1993, Cell.

[57]  K. Alitalo,et al.  Vascularization of the mouse embryo: A study of flk‐1, tek, tie, and vascular endothelial growth factor expression during development , 1995, Developmental dynamics : an official publication of the American Association of Anatomists.

[58]  A. Logan,et al.  Angiogenesis , 1993, The Lancet.

[59]  J. Aubin,et al.  Parathyroid hormone reversibly suppresses the differentiation of osteoprogenitor cells into functional osteoblasts. , 1990, Endocrinology.