Download PDF
Thromb Haemost 1990; 64(03): 412-419
DOI: 10.1055/s-0038-1647329
Original Article
Schattauer GmbH Stuttgart

Induction of Synthesis of Plasminogen Activator Inhibitor Type-1 byTissue-Type Plasminogen Activator in Human Hepatic and Endothelial Cells

Satoshi Fujii
1   The Cardiovascular Division, Washington University School of Medicine, St. Louis, MO, U.S.A.
,
Charles L Lucore
1   The Cardiovascular Division, Washington University School of Medicine, St. Louis, MO, U.S.A.
,
William E Hopkins
1   The Cardiovascular Division, Washington University School of Medicine, St. Louis, MO, U.S.A.
,
Joseph J Billadello
1   The Cardiovascular Division, Washington University School of Medicine, St. Louis, MO, U.S.A.
,
Burton E Sobel
1   The Cardiovascular Division, Washington University School of Medicine, St. Louis, MO, U.S.A.
› Author Affiliations
Further Information

Publication History

Received 14 March 1990

Accepted after revision 08 June 1990

Publication Date:
04 September 2018 (online)

Also available at
  PDF Download  Permissions and Reprints

Summary

Plasminogen activator inhibitor type-1 (PAI-1) can modify fibrinolytic activity in vitro and in vivo. The present study was performed to determine whether pharmacologic concentrations of tissue-typeplasminogen activator (t-PA) can initiate negative feedback by stimulating PAI-1 synthesis. In both human hepatoma cells (Hep G2) and human umbilical vein endothelial cells (HUVEC), t-PA increased the total concentrations and appearance of newly synthesized protein inconditioned media of free PAI-1 and PAI-1 complexed with t-PA in a dose and time dependent fashion judging from results after immunoprecipi-tation of metabolically labeled PAI-1. The t-PA effect was not attributable simply to release of stored or matrix-bound PAI-1. In HUVEC, Northern blot analyses indicated that t-PA increased steady-state levels of PAI-1 mRNA two-fold. In contrast PAI-1 mRNA expression was not increased in Hep G2 cells. Thus, mechanisms of stimulation appeared to differ in the two cell lines. The results obtained are consistent with the hypothesis that increased PAI-1 synthesis and secretion in response to t-PA may limit or attenuate fibrinolysis locally or systemically in vivo.

 

  • References

  • 1 DeWood MA, Spores J, Notske RN, Mouser LI, Burroughs R, Golden MS, Lang HT. Prevalence of total coronary artery occlusion during the early hours of transmural myocardial infarction. N Engl J Med 1980; 303: 897-902
    CrossrefPubMedGoogle Scholar
  • 2 Collen D, Topol EJ, Tiefenbrunn AJ, Gold HK, Weisfeldt ML, Sobel BE, Leinbach RC, Brinker JA, Ludbrook PA, Yasuda T, Bulkley BH, Robison AK, Hutter AM, Bell WR, Spadaro JJ, Khaw BA, Grossbard EB. Coronary thrombolysis with recombinant human tissue-type plasminogen activator; a prospective, randomized placebo-controlled trial. Circulation 1984; 70: l0l2-1017
    PubMedGoogle Scholar
  • 3 verstraete M, Bernard R, Bory M, Brower RW, Collen D, deBoro DP, Erbel R, Huhmann W, Lennane RJ, Lubsen J, Matthey D, Meyer J, Michels HR, Rutsch W, Schari M, Schmidt W, Uebis R, von Essen R. Randomized trial of intravenous recombinant human tissue-type plasminogen activator versus intravenous streptokinase in acute myocardial infarction. Lancet 1985; l: 842-847
    PubMedGoogle Scholar
  • 4 Sobel BE. Pharmacologic thrombolysis: tissue-type plasminogen activator. Circulation 1987; 76 suppl II 39-43
    PubMedGoogle Scholar
  • 5 Califf RM, Topol EJ, George BS, Boswick JM, Lee KL, Stump D, Dillon J, Abbottsmith C, Candela RJ, Kereiakes DJ, o’Neill WW, Stack RS. and the TAMI Study Group. Characteristics and outcome of patients in whom reperfusion with intravenous tissue-type plasminogen activator fails: results of Thrombolysis and Angioplasty in Myocardial Infarction (TAMI) I trial. Circulation 1988; 77: 1090-1099
    CrossrefPubMedGoogle Scholar
  • 6 Lucore CL, Fujii S, Sobel BE. Dependence of fibrinolytic activity on the concentration of free rather than total tissue-type plasminogen activator in plasma after pharmacologic administration. Circulation 1989; 79: 1204-1213
    CrossrefPubMedGoogle Scholar
  • 7 Lucore CL, Sobel BE. Interaction of tissue-type plasminogen activator with plasma inhibitors and their pharmacologic implications. Circulation 1988; 77: 660-669
    CrossrefPubMedGoogle Scholar
  • 8 Lucore CL, Fujii S, Wun T-C, Sobel BE, Billadello JJ. Regulation of the expression of type 1 plasminogen activator inhibitor in Hep G2 cells by epidermal growth factor. J Biol Chem 1988; 263: 15845-15848
    CrossrefPubMedGoogle Scholar
  • 9 Fujii S, Lucore CL, Hopkins WE, Billadello JJ, Sobel BE. Potential attenuation of fibrinolysis by growth factors released from platelets and their pharmacologic implications. Am J Cardiol 1989; 63: 1505-1511
    CrossrefPubMedGoogle Scholar
  • 10 Saksela O, Moscatelli D, Rifkin DB. The opposing effects of basic fibroblast growth factilr and transforming growth factor beta on the regulation of plasminogen activator activity in capillary endothelial cells. J Cell Biol 1987; 105: 957-963
    CrossrefPubMedGoogle Scholar
  • 11 Schleef RR, Bevilacqua MP, Sawdey M, Gimbrone MA, Loskutoff DJ. Cytokine activation of vascular endothelium. J Biol Chem 1988; 263: 5797-5803
    PubMedGoogle Scholar
  • 12 Colucci M, Paramo JA, Collen D. Generation in plasma of a fastacting inhibitor of plasminogen activator in response to endotoxin stimulation. J Clin Invest 1985; 75: 818-824
    CrossrefPubMedGoogle Scholar
  • 13 Gelehrter TD, Sznycer-Laszuk R. Thrombin induction of plasminogen activator-inhibitor in cultured human endothelial cells. J Clin Invest 1986; 77: 165-169
    CrossrefPubMedGoogle Scholar
  • 14 Knowles BB, Hoew CC, Aden DP. Human hepatocellular carcinoma cell lines secrete the major plasma proteins and hepatitis B surface antigen. Science 1980; 209: 497-499
    CrossrefPubMedGoogle Scholar
  • 15 Jaffe EA, Nachman RL, Becker CK, Minick CR. Culture of human endothelial cells derived from umbilical veins. J Clin Invest 1973; 52: 2745-2756
    CrossrefPubMedGoogle Scholar
  • 16 Mimuro J, Schleef RR, Loskutoff DJ. Extracellular matrix of cultured bovine aortic endothelial cells contains functionally active type 1 plasminogen acrivator inhibitor. Blood 1987; 70: 72l-729
    PubMedGoogle Scholar
  • 17 Kessler SW. Rapid isolation of antigens from cells with a staphylococcal protein A-antibody absorbent: parameters of the interactions of antibody antigen complexes with protein A. J Immunol 1975; 115: 1617-1624
    PubMedGoogle Scholar
  • 18 Laemmli UK. Cleavage of structural protein during assembly of the head of bacteriophage T4. Nature 1970; 227: 680-685
    CrossrefPubMedGoogle Scholar
  • 19 Loskutoff DJ, Van Mourik JA, Erickson LA, Lawrence D. Detection of an unusually stable fibrinolytic inhibitor produced by bovine endothelial cells. Proc Natl Acad Sci USA 1983; 80: 2956-2960
    CrossrefPubMedGoogle Scholar
  • 20 Bergdorf N, Nilsson T, Wallen P. An enzyme linked immunosorbent assay for determination of tissue plasminogen activator applied to patients with thromboembolic disease. Thromb Haemostas 1983; 50: 740-744
    PubMedGoogle Scholar
  • 21 Declerck PJ, Alessi M, Verstreken M, Kruithof EK O, Juhan-Vague I, Collen D. Measurement of plasminogen activator inhibitor of in biologic fluids with a murine monoclonal antibody-based enzymelinked immunosorbent assay. Blood 1988; 71: 220-225
    CrossrefPubMedGoogle Scholar
  • 22 Wun T-C, Kretzmer KK. cDNA cloning and expression in E. Coli of a plasminogen activator inhibitor (PAI) related to a PAI produced by Hep.G2 hepatoma cell. FEBS Lett 1987 71. 11-16
    PubMedGoogle Scholar
  • 23 Tso JX, Sun xH, Kao TH, Reece KA, wu R. Isolation and characterization of rat and human glyceraldehyde-3-phosphate dehydrogenase cDNAs: genomic complexity and molecular evolution of the gene. Nucleic Acid Res 1985; 13: 2485-2501
    CrossrefPubMedGoogle Scholar
  • 24 Feinberg A, Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 1983; 132: 6-13
    CrossrefPubMedGoogle Scholar
  • 25 Dillman WH, Barrieux A, Neeley wE, Conteras P. Influence of thyroid hormone on the in vitro translational activity of specific mRNAs in the rat heart. J Biol Chem 1983; 258: 7738-7745
    PubMedGoogle Scholar
  • 26 Thomas PS. Hybridiation of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci USA 1980; 77: 5201-5205
    CrossrefPubMedGoogle Scholar
  • 27 van Hinsberg vw M, Binnema D, Scheffer MA, Sprengers ED, Kooistra T, Rijken DC. Production of plasminogen activators and inhibitor by serially propagated endothelial cells from adult human blood vessels. Arteriosclerosis 1987; 7: 389-400
    PubMedGoogle Scholar
  • 28 Booyse FM, Scheinbuks J, Hua Lin P, Tiaylor M, Bruce R. Isolation and interrelationships of the multiple molecular tissue-type and urokinase-type plasminogen activator forms produced by cultured human umbilical vein endothelial cells. J Biol Chem 1988; 263: 15129-15138
    PubMedGoogle Scholar
  • 29 Wojta J, Hoover RL, Daniel TO. Vascular origin determines plasminogen activator expression in human endothelial cells. Renal endothelial cells produce large amounts of single chain urokinase type plasminogen activator J Biol Chem 1989; 264: 2846-2852
    PubMedGoogle Scholar
  • 30 van Mourik JA, Lawrence DA, Loskutoff DJ. Purification of an inhibitor of plasminogen activator synthesized by endothelial cells. J Biol Chem 1984; 259: 14914-14921
    CrossrefPubMedGoogle Scholar
  • 31 Knudsen BS, Nachman RL. Matrix plasminogen activator inhibitor. J Biol Chem 1988; 263: 9476-9481
    PubMedGoogle Scholar
  • 32 Duff Gw, Atkins E. The inhibitory effect of polymyxin B on endotoxin-induced endogenous pyrogen production. J Immunol Methods 1980; 52: 333-340
    PubMedGoogle Scholar
  • 33 Ginsburg D, Zeheb R, Yang AY, Rafferty UM, Andreasen pA, Nielsen L, Dano K, Lebo RV, Gelehrter TD. cDNA cloning of human plasminogen activator inhibitor from endothelial cells. I Ctin Invest 1986; 78: 1673-1680
    PubMedGoogle Scholar
  • 34 Tiefenbrunn AJ, Sobel BE. The impact of coronary thrombolysis on myocardial infarction. Fibrinolysis 1989; 3: 1-15
    PubMedGoogle Scholar
  • 35 Kooistra T, Sprengers ED, van Hinsbergh VW M. Rapid inactivation of the plasminogen-activator inhibitor upon secretion from cultured human endothelial cells. Biochem J 1986; 239: 497-403
    CrossrefPubMedGoogle Scholar
  • 36 Garabedian HD, Gold HK, Leinbach RC, Johns JA, yasuda T, Kanke M, Collen D. Comparative properties of two clinical preparations of recombinant human tissue-type plasminogen activator in patients with acute myocardial infarction. J Am Coll Cardiol 1987; 9: 599-607
    CrossrefPubMedGoogle Scholar
  • 37 Moore KL, Andreoli sR, Esmon NL, Esmon cI, Bang NU. Endotoxin enhances tissue factor and suppresses thrombomodulin expression of human vascular endothelium in vitro. J Clin Invest 1987; 79: 124-130
    CrossrefPubMedGoogle Scholar
  • 38 van den Berg EA, Sprengers ED, Jaye M, Burgess W, Maciag T, van Hinsbergh VW M. Regulation of plasminogen activator inhibitor-1 mRNA in human endothelial cells. Thromb Haemostas 1988; 60: 63-67
    PubMedGoogle Scholar
  • 39 Sakata Y, Okado M, Norn A, Matsuda M. Interaction of tissue-type plasminogen activator and plasminogen activator inhibitor 1 on the surface of endothelial cells. J Biol Chem 1988; 263: 1960-1969
    PubMedGoogle Scholar
  • 40 Levin EG. Quantitation and properties of the active and latent plasminogen activator inhibitors in cultures of human endothelial cells. Blood 1986; 67: 1309-1313
    PubMedGoogle Scholar
  • 41 Owensby DA, Sobel BE, Schwartz AL. Receptor-mediated endocytosis of tissue-type plasminogen activator by the human hepatoma cell line Hep G2. J Biol Chem 1988; 263:: 10587-10594
    PubMedGoogle Scholar
  • 42 Owensby DA, Morton PA, Schwartz AL. Interactions between tissue-type plasminogen activator and extracellular matrix-associated plasminogen activator inhibitor type 1 in the human hepatoma cell line Hep G2. J Biol Chem 1989; 264: 18180-18187
    PubMedGoogle Scholar
  • 43 Ord JM, Owensby DA, Billadello JJ, Sobel BR. Rapid clearance of tissue-type plasminogen activator despite diminished interaction with plasminogen activator inhibitor type . Fibrinolysis 1990; 4: 203-209
    PubMedGoogle Scholar
  • 44 Sobel BE. Coronary thrombolysis and the new biology. J Am Coll Cardiol 1989; 14: 850-860
    CrossrefPubMedGoogle Scholar