Angiogenesis: role of calcium-mediated signal transduction.
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Journal: 1995/April - Proceedings of the National Academy of Sciences of the United States of America
ISSN: 0027-8424
PUBMED: 7533291
Abstract:
During angiogenesis, endothelial cells react to stimulation with finely tuned signaling responses. The role of calcium-regulated signaling in angiogenesis has not been defined. This study investigated the calcium dependency of endothelial cell proliferation and invasion by using an inhibitor of ligand-stimulated calcium influx, CAI (carboxy-amidotriazole). Incubation with CAI significantly inhibited proliferation of human umbilical vein endothelial cells (HUVECs) in response to serum (IC50 = 1 microM) or basic fibroblast growth factor (FGF2; P2 < 0.005 at 10 microM). Statistically significant inhibition of HUVEC adhesion and motility to basement membrane proteins laminin, fibronectin, and type IV collagen was demonstrated (adhesion, P2 < 0.004-0.01; motility, P2 < 0.009-0.018). Marked inhibition of native and FGF2-induced gelatinase activity was shown by zymogram analysis and was confirmed by Northern blot analysis. CAI inhibited HUVEC tube formation on Matrigel and inhibited in vivo angiogenesis in the chicken chorioallantoic membrane assay, 67% at 20 microM and 56% at 10 microM compared with 16% for an inactive CAI analog or 9% for 0.1% dimethyl sulfoxide control. Incubation of HUVECs with CAI and/or FGF2 followed by immunoprecipitation with anti-phosphotyrosine antibody showed inhibition of FGF2-induced tyrosine phosphorylation of proteins in the range 110-150 kDa. These results suggest that calcium-regulated events are important in native and FGF2-stimulated HUVEC proliferation and invasion, perhaps through regulation of FGF2-induced phosphorylation events, and indicate a role for calcium in the regulation of angiogenesis in vivo.
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Proc Natl Acad Sci U S A 92(5): 1307-1311
PMID: 7533291

Angiogenesis: role of calcium-mediated signal transduction.

Abstract

During angiogenesis, endothelial cells react to stimulation with finely tuned signaling responses. The role of calcium-regulated signaling in angiogenesis has not been defined. This study investigated the calcium dependency of endothelial cell proliferation and invasion by using an inhibitor of ligand-stimulated calcium influx, CAI (carboxy-amidotriazole). Incubation with CAI significantly inhibited proliferation of human umbilical vein endothelial cells (HUVECs) in response to serum (IC50 = 1 microM) or basic fibroblast growth factor (FGF2; P2 < 0.005 at 10 microM). Statistically significant inhibition of HUVEC adhesion and motility to basement membrane proteins laminin, fibronectin, and type IV collagen was demonstrated (adhesion, P2 < 0.004-0.01; motility, P2 < 0.009-0.018). Marked inhibition of native and FGF2-induced gelatinase activity was shown by zymogram analysis and was confirmed by Northern blot analysis. CAI inhibited HUVEC tube formation on Matrigel and inhibited in vivo angiogenesis in the chicken chorioallantoic membrane assay, 67% at 20 microM and 56% at 10 microM compared with 16% for an inactive CAI analog or 9% for 0.1% dimethyl sulfoxide control. Incubation of HUVECs with CAI and/or FGF2 followed by immunoprecipitation with anti-phosphotyrosine antibody showed inhibition of FGF2-induced tyrosine phosphorylation of proteins in the range 110-150 kDa. These results suggest that calcium-regulated events are important in native and FGF2-stimulated HUVEC proliferation and invasion, perhaps through regulation of FGF2-induced phosphorylation events, and indicate a role for calcium in the regulation of angiogenesis in vivo.

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Selected References

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  • Folkman J, Shing Y. Angiogenesis. J Biol Chem. 1992 Jun 5;267(16):10931–10934. [PubMed] [Google Scholar]
  • Folkman J. The role of angiogenesis in tumor growth. Semin Cancer Biol. 1992 Apr;3(2):65–71. [PubMed] [Google Scholar]
  • Bouck N. Tumor angiogenesis: the role of oncogenes and tumor suppressor genes. Cancer Cells. 1990 Jun;2(6):179–185. [PubMed] [Google Scholar]
  • Folkman J, Klagsbrun M. Angiogenic factors. Science. 1987 Jan 23;235(4787):442–447. [PubMed] [Google Scholar]
  • Yarden Y, Ullrich A. Growth factor receptor tyrosine kinases. Annu Rev Biochem. 1988;57:443–478. [PubMed] [Google Scholar]
  • Meldolesi J, Clementi E, Fasolato C, Zacchetti D, Pozzan T. Ca2+ influx following receptor activation. Trends Pharmacol Sci. 1991 Aug;12(8):289–292. [PubMed] [Google Scholar]
  • Berridge MJ, Irvine RF. Inositol phosphates and cell signalling. Nature. 1989 Sep 21;341(6239):197–205. [PubMed] [Google Scholar]
  • Lin LL, Wartmann M, Lin AY, Knopf JL, Seth A, Davis RJ. cPLA2 is phosphorylated and activated by MAP kinase. Cell. 1993 Jan 29;72(2):269–278. [PubMed] [Google Scholar]
  • Gusovsky F, Lueders JE, Kohn EC, Felder CC. Muscarinic receptor-mediated tyrosine phosphorylation of phospholipase C-gamma. An alternative mechanism for cholinergic-induced phosphoinositide breakdown. J Biol Chem. 1993 Apr 15;268(11):7768–7772. [PubMed] [Google Scholar]
  • Felder CC, MacArthur L, Ma AL, Gusovsky F, Kohn EC. Tumor-suppressor function of muscarinic acetylcholine receptors is associated with activation of receptor-operated calcium influx. Proc Natl Acad Sci U S A. 1993 Mar 1;90(5):1706–1710.[PMC free article] [PubMed] [Google Scholar]
  • Moody TW, Cuttitta F. Growth factor and peptide receptors in small cell lung cancer. Life Sci. 1993;52(14):1161–1173. [PubMed] [Google Scholar]
  • Takigawa M, Nishida Y, Suzuki F, Kishi J, Yamashita K, Hayakawa T. Induction of angiogenesis in chick yolk-sac membrane by polyamines and its inhibition by tissue inhibitors of metalloproteinases (TIMP and TIMP-2). Biochem Biophys Res Commun. 1990 Sep 28;171(3):1264–1271. [PubMed] [Google Scholar]
  • Savarese DM, Russell JT, Fatatis A, Liotta LA. Type IV collagen stimulates an increase in intracellular calcium. Potential role in tumor cell motility. J Biol Chem. 1992 Oct 25;267(30):21928–21935. [PubMed] [Google Scholar]
  • Felder CC, Ma AL, Liotta LA, Kohn EC. The antiproliferative and antimetastatic compound L651582 inhibits muscarinic acetylcholine receptor-stimulated calcium influx and arachidonic acid release. J Pharmacol Exp Ther. 1991 Jun;257(3):967–971. [PubMed] [Google Scholar]
  • Kohn EC, Jacobs W, Kim YS, Alessandro R, Stetler-Stevenson WG, Liotta LA. Calcium influx modulates expression of matrix metalloproteinase-2 (72-kDa type IV collagenase, gelatinase A). J Biol Chem. 1994 Aug 26;269(34):21505–21511. [PubMed] [Google Scholar]
  • Kohn EC, Felder CC, Jacobs W, Holmes KA, Day A, Freer R, Liotta LA. Structure-function analysis of signal and growth inhibition by carboxyamido-triazole, CAI. Cancer Res. 1994 Feb 15;54(4):935–942. [PubMed] [Google Scholar]
  • Bavisotto LM, Schwartz SM, Heimark RL. Modulation of Ca2(+)-dependent intercellular adhesion in bovine aortic and human umbilical vein endothelial cells by heparin-binding growth factors. J Cell Physiol. 1990 Apr;143(1):39–51. [PubMed] [Google Scholar]
  • Minniti CP, Kohn EC, Grubb JH, Sly WS, Oh Y, Müller HL, Rosenfeld RG, Helman LJ. The insulin-like growth factor II (IGF-II)/mannose 6-phosphate receptor mediates IGF-II-induced motility in human rhabdomyosarcoma cells. J Biol Chem. 1992 May 5;267(13):9000–9004. [PubMed] [Google Scholar]
  • Brown PD, Levy AT, Margulies IM, Liotta LA, Stetler-Stevenson WG. Independent expression and cellular processing of Mr 72,000 type IV collagenase and interstitial collagenase in human tumorigenic cell lines. Cancer Res. 1990 Oct 1;50(19):6184–6191. [PubMed] [Google Scholar]
  • Taylor S, Folkman J. Protamine is an inhibitor of angiogenesis. Nature. 1982 May 27;297(5864):307–312. [PubMed] [Google Scholar]
  • Yamamoto N, Matsutani S, Yoshitake Y, Nishikawa K. Immunohistochemical localization of basic fibroblast growth factor in A431 human epidermoid carcinoma cells. Histochemistry. 1991;96(6):479–485. [PubMed] [Google Scholar]
  • Dell'Era P, Presta M, Ragnotti G. Nuclear localization of endogenous basic fibroblast growth factor in cultured endothelial cells. Exp Cell Res. 1991 Feb;192(2):505–510. [PubMed] [Google Scholar]
  • Kubota Y, Kleinman HK, Martin GR, Lawley TJ. Role of laminin and basement membrane in the morphological differentiation of human endothelial cells into capillary-like structures. J Cell Biol. 1988 Oct;107(4):1589–1598.[PMC free article] [PubMed] [Google Scholar]
  • Jaye M, Schlessinger J, Dionne CA. Fibroblast growth factor receptor tyrosine kinases: molecular analysis and signal transduction. Biochim Biophys Acta. 1992 Jun 10;1135(2):185–199. [PubMed] [Google Scholar]
  • Kohn EC, Liotta LA. L651582: a novel antiproliferative and antimetastasis agent. J Natl Cancer Inst. 1990 Jan 3;82(1):54–60. [PubMed] [Google Scholar]
  • Kohn EC, Liotta LA, Schiffmann E. Autocrine motility factor stimulates a three-fold increase in inositol trisphosphate in human melanoma cells. Biochem Biophys Res Commun. 1990 Jan 30;166(2):757–764. [PubMed] [Google Scholar]
  • Kohn EC, Sandeen MA, Liotta LA. In vivo efficacy of a novel inhibitor of selected signal transduction pathways including calcium, arachidonate, and inositol phosphates. Cancer Res. 1992 Jun 1;52(11):3208–3212. [PubMed] [Google Scholar]
  • Rifkin DB, Moscatelli D. Recent developments in the cell biology of basic fibroblast growth factor. J Cell Biol. 1989 Jul;109(1):1–6.[PMC free article] [PubMed] [Google Scholar]
  • Gospodarowicz D. Biological activities of fibroblast growth factors. Ann N Y Acad Sci. 1991;638:1–8. [PubMed] [Google Scholar]
  • Montesano R, Pepper MS, Belin D, Vassalli JD, Orci L. Induction of angiogenesis in vitro by vanadate, an inhibitor of phosphotyrosine phosphatases. J Cell Physiol. 1988 Mar;134(3):460–466. [PubMed] [Google Scholar]
  • Akiyama T, Ishida J, Nakagawa S, Ogawara H, Watanabe S, Itoh N, Shibuya M, Fukami Y. Genistein, a specific inhibitor of tyrosine-specific protein kinases. J Biol Chem. 1987 Apr 25;262(12):5592–5595. [PubMed] [Google Scholar]
  • Fotsis T, Pepper M, Adlercreutz H, Fleischmann G, Hase T, Montesano R, Schweigerer L. Genistein, a dietary-derived inhibitor of in vitro angiogenesis. Proc Natl Acad Sci U S A. 1993 Apr 1;90(7):2690–2694.[PMC free article] [PubMed] [Google Scholar]
  • Presta M, Maier JA, Ragnotti G. The mitogenic signaling pathway but not the plasminogen activator-inducing pathway of basic fibroblast growth factor is mediated through protein kinase C in fetal bovine aortic endothelial cells. J Cell Biol. 1989 Oct;109(4 Pt 1):1877–1884.[PMC free article] [PubMed] [Google Scholar]
  • Huang AJ, Manning JE, Bandak TM, Ratau MC, Hanser KR, Silverstein SC. Endothelial cell cytosolic free calcium regulates neutrophil migration across monolayers of endothelial cells. J Cell Biol. 1993 Mar;120(6):1371–1380.[PMC free article] [PubMed] [Google Scholar]
  • Fafeur V, Jiang ZP, Böhlen P. Signal transduction by bFGF, but not TGF beta 1, involves arachidonic acid metabolism in endothelial cells. J Cell Physiol. 1991 Nov;149(2):277–283. [PubMed] [Google Scholar]
Signal Transduction and Prevention Unit, National Cancer Institute, Bethesda, MD 20892.
Signal Transduction and Prevention Unit, National Cancer Institute, Bethesda, MD 20892.
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Abstract
During angiogenesis, endothelial cells react to stimulation with finely tuned signaling responses. The role of calcium-regulated signaling in angiogenesis has not been defined. This study investigated the calcium dependency of endothelial cell proliferation and invasion by using an inhibitor of ligand-stimulated calcium influx, CAI (carboxy-amidotriazole). Incubation with CAI significantly inhibited proliferation of human umbilical vein endothelial cells (HUVECs) in response to serum (IC50 = 1 microM) or basic fibroblast growth factor (FGF2; P2 < 0.005 at 10 microM). Statistically significant inhibition of HUVEC adhesion and motility to basement membrane proteins laminin, fibronectin, and type IV collagen was demonstrated (adhesion, P2 < 0.004-0.01; motility, P2 < 0.009-0.018). Marked inhibition of native and FGF2-induced gelatinase activity was shown by zymogram analysis and was confirmed by Northern blot analysis. CAI inhibited HUVEC tube formation on Matrigel and inhibited in vivo angiogenesis in the chicken chorioallantoic membrane assay, 67% at 20 microM and 56% at 10 microM compared with 16% for an inactive CAI analog or 9% for 0.1% dimethyl sulfoxide control. Incubation of HUVECs with CAI and/or FGF2 followed by immunoprecipitation with anti-phosphotyrosine antibody showed inhibition of FGF2-induced tyrosine phosphorylation of proteins in the range 110-150 kDa. These results suggest that calcium-regulated events are important in native and FGF2-stimulated HUVEC proliferation and invasion, perhaps through regulation of FGF2-induced phosphorylation events, and indicate a role for calcium in the regulation of angiogenesis in vivo.
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