Download PDF
Thromb Haemost 1988; 60(01): 063-067
DOI: 10.1055/s-0038-1647636
Original Article
Schattauer GmbH Stuttgart

Regulation of Plasminogen Activator Inhibitor-1 mRNA in Human Endothelial Cells

E A van den Berg
1   The Gaubius Institute TNO, Leiden, The Netherlands
,
E D Sprengers
1   The Gaubius Institute TNO, Leiden, The Netherlands
,
M Jaye
2   The Rorer Biotechnology Inc., Springfield, VA, USA
,
W Burgess
3   The Jerome H. Holland Laboratory for the Biomedical Sciences, American Red Cross, Rockville, MD, USA
,
T Maciag
3   The Jerome H. Holland Laboratory for the Biomedical Sciences, American Red Cross, Rockville, MD, USA
,
V W M van Hinsbergh
1   The Gaubius Institute TNO, Leiden, The Netherlands
› Author Affiliations
Further Information

Publication History

Received 02 November 1987

Accepted after revision 05 April 1988

Publication Date:
30 June 2018 (online)

  PDF Download  Permissions and Reprints

Summary

The plasminogen activator inhibitor (PAI-1) from endothelial cells is a potentially important regulator of plasminogen activator activity. Cultured human endothelial cells increase their PAI-1 production upon stimulation with LPS and TNF, agents that are known to cause an increase in PAI-1 levels in vivo.

We isolated a PAI-1 cDNA probe, and by RNA hybridization analysis studied the regulation of PAI-1 mRNA synthesis in human endothelial artery cells. Freshly isolated endothelial cells do not contain detectable amounts of PAI-1 mRNA, but after adherence and incubation for 18 h in growth medium produce considerable amounts of PAI-1 activity and contain PAI-1 mRNA levels comparable to those found in subcultured cells. When subcultured endothelial cells are incubated for 6 h with LPS or TNF, both species of PAI-1 mRNA increase 10 to 20 fold, while PAI-1 activity in the growth medium increases only 1.5 to 2 fold. Stimulation of endothelial cells in the presence of cycloheximide (CHX) results in superinduction of mainly the 3.0 kb PAI-1 mRNA. The 3' end of this mRNA contains a 60 bp AT-rich sequence, that resembles 3' sequences present in a number of other genes superinducible with CHX.

Key words

Endothelial cells - Plasminogen-activator inhibitor - Messenger RNA - Tumor necrosis factor - Endotoxin
 

  • References

  • 1 Collen D. On the regulation and control of fibrinolysis. Thromb Haemostas 1980; 43: 77-89
    PubMedGoogle Scholar
  • 2 Erickson LA, Schleef RR, Ny T, Loskutoff DJ. The fibrinolytic system of the vascular wall. Clin Haematol 1985; 14: 513-530
    PubMedGoogle Scholar
  • 3 Danø K, Andreasen PA, Grøndahl-Hansen J, Kristensen P, Nielsen LS, Skriver L. Plasminogen activators, tissue degradation and cancer. Adv Cancer Res 1985; 44: 140-239
    PubMedGoogle Scholar
  • 4 Sprengers ED, Kluft C. Plasminogen activator inhibitors. Blood 1987; 69: 381-387
    PubMedGoogle Scholar
  • 5 Kruithof EK O, Tran-Thang C, Gudinchet A, Hauert J, Nicoloso G, Genton C, Welti H. Bachmann E Fibrinolysis in pregnancy: a study of plasminogen activator inhibitors. Blood 1987; 69: 460-466
    PubMedGoogle Scholar
  • 6 Chapman Jr HA, Stone OL. Characterization of a macrophage- derived plasminogen activator inhibitor. Biochem J 1985; 230: 109-116
    CrossrefPubMedGoogle Scholar
  • 7 Juhan-Vague I, Moerman B, Decock F, Aillaud MF, Collen D. Plasma levels of a specific inhibitor of tissue type plasminogen, activator (and urokinase) in normal and pathological conditions. Thromb Res 1984; 33: 523-530
    CrossrefPubMedGoogle Scholar
  • 8 Colucci M, Paramo JA, Collen D. Generation in plasma of a fast acting inhibitor of plasminogen activator in response to endotoxin stimulation. J Clin Invest 1985; 75: 818-824
    CrossrefPubMedGoogle Scholar
  • 9 Hamsten A, Wiman B, De Faire U, Blomback M. Increased plasma levels of a rapid inhibitor of tissue plasminogen activator in young survivors of myocardial infarction. New Engl J Med 1985; 313: 1557-1563
    CrossrefPubMedGoogle Scholar
  • 10 Kluft C, Jie AF H, Lowe GD O, Blamey SL, Forbes CD. Association between postoperative hyperresponse in t-PA inhibition and deep vein thrombosis. Thromb Haemostas 1986; 56: 107
    PubMedGoogle Scholar
  • 11 Emeis JJ, Kooistra T. Interleukin 1 and lipopolysaccharide induce an inhibitor of tissue type plasminogen activator in vivo and in cultured endothelial cells. J Exp Med 1986; 163: 1260-1266
    CrossrefPubMedGoogle Scholar
  • 12 Van Hinsbergh VW M, Kooistra T, Fiers W, Emeis JJ. Tumor necrosis factor increases the production of plasminogen activator inhibitor in human endothelial cells in vitro and rats in vivo. Thromb Haemostas 1987; 58: 15
    PubMedGoogle Scholar
  • 13 Marmenout A, Fransen L, Tavernier J, Van der Hayden J, Tizard R, Kawashima E, Shaw A, Johnson MJ, Semon D, Muller R, Ruys-schaert MR, Van Vliet A, Fiers W. Molecular cloning and expression of human tumor necrosis factor and comparison with mouse tumor necrosis factor. Eur J Biochem 1985; 152: 512-519
    PubMedGoogle Scholar
  • 14 Doodemont H, Soriano P, Quax WJ, Ramaekers F, Leustra JA, Groenen MAM, Bernardi G, Bloemendaal H. The genes coding for the cytoskeletal proteins actin and vimentin in warm blooded vertebrates. EMBO J 1982; 1: 167-171
    PubMedGoogle Scholar
  • 15 Tong BD, Levine SE, Jaye M, Ricca G, Drohan W, Maciag T, Deuel TF. Isolation and sequencing of a cDNA clone homologous to the v-sis oncogen from human endothelial cells. Mol Cell Biol 1986; 6: 3018-3022
    CrossrefPubMedGoogle Scholar
  • 16 Sprengers ED, Van Hinsbergh VW M, Jansen B. The active and the inactive plasminogen activator inhibitor from human endothelial cell conditioned medium are immunologically and functionally related to each other. Biochim Biophys Acta 1986; 883: 233-241
    CrossrefPubMedGoogle Scholar
  • 17 Huyinh TV, Young RA, Davis RW. Construction and screening of cDNA libraries in lambda gt10 and gt11. In: DNA Cloning. Glover D. (ed) IRL Press; Oxford - Washington DC: 1985. 1 49-78
    Google Scholar
  • 18 Maniatis T, Fritsch EF, Sambrook J. Molecular Cloning; A Laboratory Manual. Cold Spring Harbour Laboratory: 1982. ISBN 0-87969136-0
    Google Scholar
  • 19 Sanger F, Coulson AR, Barrell BG, Smith AJ H, Roe B. Cloning in single stranded bacteriophage as an aid to rapid DNA sequencing. J Mol Biol 1980; 143: 161
    CrossrefPubMedGoogle Scholar
  • 20 Rijken DC, Van Hinsbergh VW M, Sens EH C. Quantitation of tissue-type plasminogen activator in human endothelial cell cultures by use of an enzyme immunoassay. Thromb Res 1984; 33: 145-153
    CrossrefPubMedGoogle Scholar
  • 21 Ny T, Sawdey M, Lawrence D, Millan JL, Loskutoff DJ. Cloning and sequencing of a cDNA coding for the human B-migrating endothelial cell type plasminogen activator inhibitor. Proc Natl Acad Sci USA 1986; 83: 6776-6780
    CrossrefPubMedGoogle Scholar
  • 22 Pannekoek H, Veerman H, Lambers H, Diergaarde P, Verweij CL, Van Zonneveld AJ, Van Mourik JA. Endothelial plasminogen activator inhibitor (PAI): a new member of the serpin family. EMBO J 1986; 5: 2539-2544
    PubMedGoogle Scholar
  • 23 Ginsburg D, Zeheb R, Young AY, Rafferty UM, Andreasen PA, Nielsen L, Danø K, Lebo RV, Gelehrter TD. cDNA cloning of human plasminogen activator inhibitor from endothelial cells. J Clin Invest 1987; 78: 1673-1680
    PubMedGoogle Scholar
  • 24 Lizardi PM, Engelberg A. Rapid isolation of RNA using proteinase K and sodium perchlorate. Anal Biochem 1979; 98: 116-122
    CrossrefPubMedGoogle Scholar
  • 25 Church GM, Gilbert W. Genomic sequencing. Proc Natl Acad Sci USA 1984; 81: 1991-1995
    CrossrefPubMedGoogle Scholar
  • 26 Verheijen JH, Chang GT G, Kluft C. Evidence for the occurrence of a fast-acting inhibitor for tissue-type plasminogen activator in human plasma. Thromb Haemostas 1984; 51: 392-395
    PubMedGoogle Scholar
  • 27 Levin EG, Santell L. Conversion of active to latent plasminogen activator inhibitor from human endothelial cells. Blood 1987; 70: 1090-1098
    PubMedGoogle Scholar
  • 28 Hekman CM, Loskutoff OJ. Endothelial cells produce a latent inhibitor of plasminogen activators that can be activated by denaturants. J Biol Chem 1985; 260: 11581-11587
    PubMedGoogle Scholar
  • 29 Jaffe EA, Nachman RL, Becker CG, Minick CR. Culture of human endothelial cells derived from umbilical veins, identification by morphologic and immunologic criteria. J Clin Invest 1973; 52: 2745-2756
    CrossrefPubMedGoogle Scholar
  • 30 Maciag T, Cerundolo J, Ilsley S, Kelley PR, Forand R. An endothelial cell growth factor from bovine hypothalamus: identification and partial characterization. Proc Natl Acad Sci USA 1979; 76: 5674-5678
    CrossrefPubMedGoogle Scholar
  • 31 Van Hinsbergh VW M, Bertina RM, Van Wijngaarden A, Van Tilburg NH, Emeis JJ, Haverkate F. Activated protein C decreases plasminogen activator inhibitor activity in endothelial cell conditioned medium. Blood 1985; 65: 444-451
    PubMedGoogle Scholar
  • 32 Pollarien J, Saksela O, Salonen E, Andreasen P, Nielsen L. Danø K, Vaheri A. Distinct localization of urokinase-type plasminogen activator and its type I inhibitor under cultured fibroblasts and sarcoma cells. J Cell Biol 1987; 104: 1085-1096
    CrossrefPubMedGoogle Scholar
  • 33 Rheinwald JG, Jorgensen JL, Hahm WC, Terpstra AJ, O’Connell TM, Plummer KK. Mesosecrin: a secreted glycoprotein produced in abundance by human mesothelial, endothelial and kidney epithelial cells in culture. J Cell Biol 1987; 104: 263-275
    CrossrefPubMedGoogle Scholar
  • 34 Chapman Jr HA, Vavrin Z, Hibbs Jr JB. Macrophage fibrinolytic activity: identification of two pathways of plasmin formation by intact cells and of a plasminogen activator inhibitor. Cell 1982; 28: 653-662
    CrossrefPubMedGoogle Scholar
  • 35 Webb AC, Collins KL, Snyder SE, Alexander SJ, Rosenwasser LJ, Eddy RL, Shows TB, Auron PE. Human monocyte ARG-serpin cDNA: sequence, chromocomal assignment and homology to plasminogen activator inhibitor. J Exp Med 1987; 166: 77-94
    CrossrefPubMedGoogle Scholar
  • 36 Movat HZ, Cybulski MI, Colditz IG, Chan MK W, Dinarello CA. Acute inflammation in gram negative infection: endotoxin, interleukin 1, tumor necrosis factor and neutrophils. Fed Proc 1987; 46: 97-104
    PubMedGoogle Scholar
  • 37 Gamble JR, Harlan JM, Klebanoff SJ, Vadas MA. Stimulation of the adherence of neutrophils to umbilical vein endothelium by human recombinant tumor necrosis factor. Proc Natl Acad Sci USA 1985; 82: 8867-8871
    PubMedGoogle Scholar
  • 38 Collins T, Lapierre LA, Fiers W, Strominger JL, Pober JS. Recombinant human tumor necrosis factor increases mRNA levels and surface expression of HLA-A,B antigens in vascular endothelial cells and dermal fibroblasts in vitro. Proc Natl Acad Sci USA 1986; 83: 446-450
    CrossrefPubMedGoogle Scholar
  • 39 Broudy VC, Kaushansky K, Segal GM, Harlan JM, Adamson JW. Tumor necrosis factor type alpha stimulates human endothelial cells to produce granulocyte/macrophage colony stimulating factor. Proc Natl Acad Sci USA 1986; 83: 7467-7471
    CrossrefPubMedGoogle Scholar
  • 40 Lund LR, Riccio A, Andreasen PA, Nielsen LS, Kristensen P, Laiho M, Saksela O, Blasi F, Danø K. Transforming growth factor-ß is a strong and fast acting positive regulator of the level of type-1 plasminogen activator inhibitor mRNA in WI-38 human lung fibroblasts. EMBO J 1987; 6: 1281-1286
    PubMedGoogle Scholar
  • 41 Collart MA, Belin D, Vassalli J, de Kossodo S, Vassalli P. Gamma interferon macrophage transcription of the tumor necrosis factor/ cachectin, interleukin 1 and urokinase genes, which are controlled by short lived repressors. J Exp Med 1986; 164: 2113-2118
    CrossrefPubMedGoogle Scholar
  • 42 Mitchell RL, Henning-Chubb C, Hubermann E, Verma IM. C-fos expression is neither sufficient nor obligatory for differentiation of monomyelocytes to macrophages. Cell 1986; 45: 497-504
    CrossrefPubMedGoogle Scholar
  • 43 Linial M, Gunderson N, Groudine M. Enhanced transcription of c-myc in bursal lymphoma cells requires continuous protein synthesis. Science 1985; 230: 1126-1132
    CrossrefPubMedGoogle Scholar
  • 44 Thorens B, Mermod JJ, Vassali P. Phagocytosis and inflammatory stimuli induce GM-CSF mRNA in macrophages through posttranscriptional regulation. Cell 1987; 48: 671-679
    CrossrefPubMedGoogle Scholar
  • 45 Shaw G, Kamen R. A conserved AU sequence from the 3’ untranslated region of GM-CSFmRNA mediates selective mRNA degradation. Cell 1986; 46: 659-667
    CrossrefPubMedGoogle Scholar
  • 46 Caput D, Beutler B, Hartog K, Thayer R, Brown-Shimer S, Cerami A. Identification of a common nucleotide sequence in the 3’ untranslated region of mRNA molecules specifying inflammatory mediators. Proc Natl Acad Sci USA 1986; 83: 1670-1674
    CrossrefPubMedGoogle Scholar
  • 47 Brawerman GG. Determinants of mRNA stability. Cell 1987; 48: 5-6
    CrossrefPubMedGoogle Scholar
  • 48 Medcalf RL, Van den Berg EA, Schleuning WD. Glucocorticoid-modulated gene expression of tissue- and urinary-type plasminogen activator and plasminogen activator inhibitor 1 and 2. J Cell Biol 1988; 106: 971-978
    CrossrefPubMedGoogle Scholar