Aortic smooth muscle cells express and secrete vascular endothelial growth factor.

We examined whether cultured bovine aortic smooth muscle (ASM) cells express VEGF. RNA blot analysis of total cellular RNA derived from ASM cells demonstrates the expression of the VEGF gene. ASM cells release in the medium a VEGF-like endothelial cell mitogen which binds to heparin-sepharose and has an apparent molecular weight of 40-45 kDa as assessed by an HPLC gel filtration column. Consistent with VEGF, this mitogen does not stimulate the proliferation of ASM cells. Immunoblot analysis of the bioactive material with an antibody specific for VEGF demonstrates the presence of a major immunoreactive band with an apparent molecular mass of 23 kDa and a minor band with a molecular mass of approximately 18 kDa, in reducing conditions. The major band has very similar apparent molecular weight as the 165 amino-acid species of human recombinant VEGF of folliculo-stellate cells derived VEGF. These data demonstrate the expression and synthesis of VEGF by cultured ASM cells and suggest that the 164 amino-acid species is the predominant molecular form of the growth factor secreted by such cells. VEGF released by ASM cells may play a paracrine role in the maintenance of the integrity of the endothelial lining or in the abnormal proliferation of the vasa vasorum which takes place in atherosclerosis.

[1]  P. Chuang,et al.  Macromolecular transport across arterial and venous endothelium in rats. Studies with Evans blue-albumin and horseradish peroxidase. , 1990, Arteriosclerosis.

[2]  Stephen M. Schwartz,et al.  Developmentally regulated production of platelet-derived growth factor-like molecules , 1984, Nature.

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

[4]  R. Ross The pathogenesis of atherosclerosis--an update. , 1986, The New England journal of medicine.

[5]  H. Dvorak,et al.  A highly conserved vascular permeability factor secreted by a variety of human and rodent tumor cell lines. , 1986, Cancer research.

[6]  J. Fiddes,et al.  Vascular endothelial growth factor: a new member of the platelet-derived growth factor gene family. , 1989, Biochemical and biophysical research communications.

[7]  C. Heldin,et al.  Arterial smooth muscle cells express platelet-derived growth factor (PDGF) A chain mRNA, secrete a PDGF-like mitogen, and bind exogenous PDGF in a phenotype- and growth state-dependent manner , 1988, The Journal of cell biology.

[8]  D. Gospodarowicz,et al.  Isolation and characterization of a newly identified endothelial cell mitogen produced by AtT‐20 cells. , 1989, The EMBO journal.

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

[10]  D. Gospodarowicz,et al.  Structural characterization and biological functions of fibroblast growth factor. , 1987, Endocrine reviews.

[11]  J. Fiddes,et al.  Nucleotide sequence of a bovine clone encoding the angiogenic protein, basic fibroblast growth factor. , 1986, Science.

[12]  D. Gospodarowicz,et al.  Basic fibroblast growth factor: expression in cultured bovine vascular smooth muscle cells. , 1988, European journal of cell biology.

[13]  D. Goeddel,et al.  Vascular endothelial growth factor is a secreted angiogenic mitogen. , 1989, Science.

[14]  S. Weinbaum,et al.  Enhanced macromolecular permeability of aortic endothelial cells in association with mitosis. , 1988, Atherosclerosis.

[15]  S. O’Brien,et al.  Human endothelial cell growth factor: cloning, nucleotide sequence, and chromosome localization. , 1986, Science.

[16]  S. Weinbaum,et al.  Transendothelial Transport of Low Density Lipoprotein in Association with Cell Mitosis in Rat Aorta , 1989, Arteriosclerosis.

[17]  A. Barger,et al.  Hypothesis: vasa vasorum and neovascularization of human coronary arteries. A possible role in the pathophysiology of atherosclerosis. , 1984, The New England journal of medicine.

[18]  J. Winkles,et al.  Human vascular smooth muscle cells both express and respond to heparin-binding growth factor I (endothelial cell growth factor). , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[19]  P. Dicorleto,et al.  Cultured endothelial cells produce a platelet-derived growth factor-like protein. , 1983, Proceedings of the National Academy of Sciences of the United States of America.

[20]  D. Rifkin,et al.  Both normal and tumor cells produce basic fibroblast growth factor , 1986, Journal of cellular physiology.

[21]  J. Folkman,et al.  Expression of acidic and basic fibroblast growth factors in human and bovine vascular smooth muscle cells. , 1990, Growth factors.

[22]  J. Folkman,et al.  Conditioned medium from mouse sarcoma 180 cells contains vascular endothelial growth factor. , 1990, Growth factors.

[23]  R. Ross,et al.  The biology of platelet-derived growth factor , 1986, Cell.

[24]  W. Rutter,et al.  Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. , 1979, Biochemistry.

[25]  J. Folkman,et al.  Endothelial cell-derived basic fibroblast growth factor: synthesis and deposition into subendothelial extracellular matrix. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[26]  R. Ma A reassessment of endothelial injury and arterial lesion formation. , 1985 .

[27]  J. Folkman,et al.  Long-term culture of capillary endothelial cells. , 1979, Proceedings of the National Academy of Sciences of the United States of America.

[28]  S. Schwartz,et al.  Replication of smooth muscle cells in vascular disease. , 1986, Circulation research.

[29]  M. Cygler,et al.  Recognition of a cell-surface oligosaccharide of pathogenic Salmonella by an antibody Fab fragment. , 1991, Science.

[30]  C. Heldin,et al.  Arterial smooth muscle cells in primary culture produce a platelet-derived growth factor-like protein. , 1985, Proceedings of the National Academy of Sciences of the United States of America.

[31]  W. N. Burnette,et al.  "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. , 1981, Analytical biochemistry.

[32]  R. Ma Endothelial regeneration. VIII. Interaction of smooth muscle cells with endothelial regrowth. , 1988 .

[33]  S. Schwartz,et al.  The role of membrane-membrane interactions in the regulation of endothelial cell growth , 1985, The Journal of cell biology.

[34]  N. Ferrara,et al.  Pituitary follicular cells secrete a novel heparin-binding growth factor specific for vascular endothelial cells. , 1989, Biochemical and biophysical research communications.

[35]  G. Conn,et al.  Amino acid and cDNA sequences of a vascular endothelial cell mitogen that is homologous to platelet-derived growth factor. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[36]  H. Phillips,et al.  Vascular endothelial growth factor is expressed in rat corpus luteum. , 1990, Endocrinology.

[37]  J. Fiddes,et al.  Capillary endothelial cells express basic fibroblast growth factor, a mitogen that promotes their own growth , 1987, Nature.

[38]  K. Miyazono,et al.  Identification of angiogenic activity and the cloning and expression of platelet-derived endothelial cell growth factor , 1989, Nature.