Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
권호 표지
DOI QR코드

DOI QR Code

Effect of Fibroblast Growth Factor-2 on the Sprouting in Vascular Endothelial Cells

혈관내피세포의 발아에 미치는 fibroblast growth factor-2의 효과

  • 김환규 (전북대학교 자연과학대학 생물과학부)
  • Published : 2004.04.01
Download PDF
⟨ Previous Next ⟩

Abstract

The sprouting of vascular endothelial cells is an initial step in angiogenesis. Matrix metalloproteinases can associate with integrin on the surface of endothelial cells, thereby promoting angiogenesis. The purpose of this study was to test if fibroblast growth factor-2 (FGF-2) can regulate the sprouting in porcine pulmonary artery endothelial cells. FGF-2 induced sprouting and secretion of MMP-2 and plasmin. FGF-2 also induced the expression of integrin Mac-1, which is inhibited IS20I. Addition of BB-94, a 2-antiplasmin and IS20I inhibited FGF-2-induced sprouting activity. Therefore, FGF-2-induced sprouting activity in PPAECs may be accomplished by secretion of proteinases such as MMP-2 and plasmin and integrin expression.

본 연구에서는 FGF-2를 이용하여 혈관내피세포의 발아와 단백질분해효소의 분비 및 인테그린과의 관계를 관찰하여 다음과 같은 결과를 얻었다. PPAECs 세포의 발아에 미치는 FGF-2의 효과를 조사한 결과, 10 ng/ml에서 약 3.5배 증가되는 등 농도-의존적으로 발아가 증가되었다. MMPs에 대한 enzyme immunoassay결과, MMP-2에서만 약 1.9배의 분비증가가 유도되었다. MMP-1 및 MMP-3는 FGF-2에 의해 유의할만한 분비 증가가 나타나지 않았으며, 특이하게 MMP-3는 FGF-2의 처리 없이도 많은 양이 분비되었고, MMP-9은 0.6∼1.2 ng/$10^{6}$ cells 정도로 분비량이 적었다. FGF-2에 의한 플라스민의 분비증가 여부를 확인한 결과, 10 ng/ml에서 약 2.6배 증가하였으며, MMPs 억제제 및 인테그린 억제제에 의해 유의할만한 감소가 나타났고 플라스민 억제제인 $\alpha$2-antiplasmin에 의해서는 플라스민의 분비가 완전히 억제되었다. 또한, FGF-2 처리에 의해 농도-의존적으로 인테그린 Mac-1의 발현이 증가되었으며, 인테그린 억제제인 IS201를 전처리한 결과, 인테그린 Mac-1의 발현이 완전히 억제되었다. FGF-2에 의한 발아 유도효과는 IS201 처리에 의해 거의 완벽하게 억제되었으며, MMP-2 및 플라스민의 분비도 유의할만하게 감소시켰다. 이상의 결과를 요약하면, FGF-2에 의해 유도된 혈관내피세포의 발아 증가는 MMP-2 및 플라스민의 분비증가와 인테그린 Mac-1의 발현 증가에 의한 것이라 생각된다.

Keywords

References

  1. Br. J. Dematol. v.131 Gelatinase activity during wound healing Agren,M.S. https://doi.org/10.1111/j.1365-2133.1994.tb04974.x
  2. Matrix Metalloproteinase 72kDa gelatinase (gelatinase A): structure, activation regulation, and substrate specificity Anita,E.Y.;A.N.Murphy;W.G.Stetler-Stevenson;Parks,W.C.(ed.);R.P.Mecham(ed.)
  3. J. Biol. Chem. v.273 Intact vitronectin induces matrix metalloproteinase-2 and tissue inhibitor of metalloproteinases-2 expression and enhanced cellular invasion by melanoma cells Bafetti,L.M.;T.N.Young;Y.Itoh;M.S.Stack https://doi.org/10.1074/jbc.273.1.143
  4. Adv. Cancer Res. v.59 The FGF family of growth factors and oncogenes Basilico,C.;D.Moscatelli https://doi.org/10.1016/S0065-230X(08)60305-X
  5. Neurosurgery v.52 IS20I, a specific alphavbeta3 integrin inhibitor, reduces glioma growth in vivo Bello,L.;V.Lucini;C.Giussani;G.Carrabba;M.Pluderi;F.Scaglione;G.Tomei;R.Villani;P.M.Black;A.Bikfalvi;R.S.Carroll https://doi.org/10.1097/00006123-200301000-00023
  6. Nat. Cell Biol. v.2 Matrix metalloproteinase-9 triggers the angiogenic switch during carcinogenesis Bergers,G.;R.Brekken;G.McMahon;T.H.Vu;T.Itoh;K.Tamaki;K.Tanzawa;P.Thorpe;S.Itohara;Z.Werbm;D.Hanahanm https://doi.org/10.1038/35036374
  7. Leukemia v.8 Angiogenic factors are hematopoietic growth factors and vice versa Bikfalvi,A;Z.C.Han
  8. Curr. Opin. Cell Biol. v.7 Proteolytic remodeling of extracellular matrix Birkedal-Hansen,H. https://doi.org/10.1016/0955-0674(95)80116-2
  9. Crit. Rev. Oral Med. v.4 Matrix metalloproteinase Birkedal-Hansan,H.;W.G.I.Moore;M.K.Boden
  10. Cell v.85 Localization of matrix metalloproteinase MMP-2 to the surface of invasive cells by interaction with integrin alpha v beta 3 Brooks,P.C.;S.Stromblad;L.C.Sanders;T.L. von Schalscha;R.T.Aimes;W.G.Stetler-Stevenson;J.P.Quigley;D.A.Cheresh https://doi.org/10.1016/S0092-8674(00)81235-0
  11. J. Biol. Chem. v.267 Angiogenesis Folkman,J.;Y.Shing
  12. Matrix Metalloproteinase The matrix metalloproteinase family Frederick,J.;J.F.Jr. Woessner;Parks,W.C.(ed.);R.P.Mecham(ed.)
  13. Science v.277 Induction of cell migration by matrix metalloprotease-2 cleavage of laminin-5 Giannelli,G.;J.Falk-Marzillier;O.Schiraldi;W.G.Stetler-Stevenson;V.Quaranta https://doi.org/10.1126/science.277.5323.225
  14. EXS v.79 Regulation of capillary formation by laminin and other components of the extracellular matrix Grant,D.S.;H.K.Kleinman
  15. Curr. Opin. Hematol v.2 Changing concepts in fibrinolysis Hajjar,K.A. https://doi.org/10.1097/00062752-199502050-00004
  16. J. Cell Biol. v.128 High affinity immunoreactive FGF receptors in the extracellular matrix of vascular endothelial cells implications for the modulation of FGF-2 Hanneken,A.;P.A.Maher;A.Baird https://doi.org/10.1083/jcb.128.6.1221
  17. J. Immunol. v.156 Bacterial lipopolysaccharide induces the association and coordinate activation of p53/56lyn and phosphatidylinositol 3-kinase in human monocytes Herrera-Velit,P.;N.E.Reiner
  18. J. Invest. Dematol. v.115 Coexpression of integrin alpha(v)beta3 and matrix metalloproteinase-2 (MMP-2) coincides with MMP-2 activation: correlation with melanoma progression Hofmann,Y.B.;J.R.Westphal;E.T.Waas;J.C.Becker;D.J.Rulter;G.N. van Muijen https://doi.org/10.1046/j.1523-1747.2000.00114.x
  19. Circ. Res. v.86 Angiopoietin-1 induces endothelial cell sprouting through the activation of focal adhesion kinase and plasmin secretion Kim I.;H.G.Kim;S.O.Moon;S.W.Chae;J.N.So;K.N.Koh;B.C.Ahn;G.Y.Koh https://doi.org/10.1161/01.RES.86.9.952
  20. J. Biol. Chem. v.274 Molecular cloning, expression, and characterization of angiopoietinrelated protein induces endothelial cell sprouting Kim,I.;S.O.Moon;K.N.Koh;H.Kim;C.S.Uhm;H.J.Kwak;N.G.Kim;G.Y.Koh https://doi.org/10.1074/jbc.274.37.26523
  21. Biochem. Biophys. Res. Commu. v.269 Lipopolysaccharide activates matrix metalloproteinase-2 in endothelial cells through an NF-kB-dependent pathway Kim,H.G.;G.Y.Koh https://doi.org/10.1006/bbrc.2000.2308
  22. Ann. Rev. Physiol. v.53 Regulators of angiogenesis Klagsbum,M.;P.D'Amore https://doi.org/10.1146/annurev.ph.53.030191.001245
  23. Mol. Biol. Cell v.4 Basic fibriblast growth factor modulates integrin expression in microvascular endothelial cells Klein,S.;F.G.Giancotti;M.Presta;S.A.Albelda;C.A.Buck;D.B.Rifkin https://doi.org/10.1091/mbc.4.10.973
  24. Microvas. Res. v.55 Vascular endothelial growth factor increase release of gelatinase A and decrease of tissue inhibitor of metalloproteinases by microvascular endothelial cells in vitro Lamoreaux,W.J.;M.E.C.Fitzgerald;A.Reiner;K.A.Hasty;S.T.Chares https://doi.org/10.1006/mvre.1997.2056
  25. Cell v.64 Cancer metastasis and angiogenesis: An imbalance of positive and negative regulation Liotta,L.A.;P.S.Steeg;W.G.Stetler-Stevenson https://doi.org/10.1016/0092-8674(91)90642-C
  26. BioAssays v.14 The matrix-degrading metalloproteinases Matrisian,L.M. https://doi.org/10.1002/bies.950140705
  27. Physiol. Rev. v.73 Biology and biochemistry of proteinases in tumor invasion Mignatti,P.;D.B.Rifkin
  28. Cell v.72 High affinity VEGF binding and developmental expression suggest Flk-1 as a major regulator of vasculogenesis and angiogenesis Millauer,B.;S.Wizigmann-Voos;H.Schnurch;R.Martinez;N.P.Moller;W.Risau;A.Ullrich https://doi.org/10.1016/0092-8674(93)90573-9
  29. J. Environ. Pathol. Toxicol. Oncol. v.22 Binding of alpha5 monoclonal antibody to cell surface alpha5beta1 integrin modulates MMP-2 and MMP-7 activity in B16F10 melanoma cells Mitra,A.;J.Chakrabarti;A.Chatterjee https://doi.org/10.1615/JEnvPathToxOncol.v22.i3.20
  30. Eur. J. Clin. Invest v.22 Regulation of angiogenesis in vitro Montesano,R. https://doi.org/10.1111/j.1365-2362.1992.tb01498.x
  31. J. Neurosci. Res. v.34 Basic fibroblast growth factor expression is required for clonogenic growth of human glioma cells Morrison,R.S.;S.Giordano;F.Yamaguchi;S.Hendrickson;M.S.Berger;K.Palczewski https://doi.org/10.1002/jnr.490340503
  32. Nat. Rev. Cancer v.2 Strategies for MMP inhibition in cancer: innovations for the post-trial era Overall,C.M.;C.Lopez-Otin https://doi.org/10.1038/nrc884
  33. Thromb. Haemost. v.86 Extracellular proteolysis and angiogenesis Pepper,M.S.
  34. Enzyme Protein v.49 Angiogenesis: a paradigm for balanced extracellular proteolysis during cell migration and morphogenesis Pepper,M.S.;R.Montesano;S.J.Mandriota;L.Orci;J.D.Vassalli
  35. Neture v.386 Mechanisms of angiogenesis Risau,W. https://doi.org/10.1038/386671a0
  36. Proc. Natl. Acad. Sci. v.98 Disruption of matrix metalloproteinase 2 binding to integrin alpha vbeta 3 by an organic molecule inhibits angiogenesis and tumor growth in vivo Silletti,S.;T.Kessler;J.Goldberg;D.L.Boger;D.A.Cheresh https://doi.org/10.1073/pnas.011343298
  37. Ann. N. Y. Acad. Sci. v.732 The family of matrix metalloproteinase Woessner,J.F.L. Jr. https://doi.org/10.1111/j.1749-6632.1994.tb24720.x
  38. I. Proc. Natl. Acad. Sci. v.97 Impaired endochondral ossification and angiogenesis in mice deficient in membrane-type matrix metalloproteinase Zhou,Z.;S.S.Apte;R.Soininen;R.Cao;G.Y.Baaklini;R.W.Rauser;J.Wang;Y.Cao;K.Tryggvason https://doi.org/10.1073/pnas.060037197