PAI-1 Synthesis in the Human Hepatoma Cell Line Hep G2 Is Increased by Cytokines - Evidence that the Liver Contributes to Acute Phase Behaviour of PAI-1

J P de Boer; J J Abbink; M C Brouwer; C Meijer; D Roem; G P Voorn; J W J Lambers; J A van Mourik; C E Hack

doi:10.1055/s-0038-1647480

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 1991; 65(02): 181-185
DOI: 10.1055/s-0038-1647480

Original Article

Schattauer GmbH Stuttgart

PAI-1 Synthesis in the Human Hepatoma Cell Line Hep G2 Is Increased by Cytokines - Evidence that the Liver Contributes to Acute Phase Behaviour of PAI-1

J P de Boer

The Central Laboratory of the Netherlands Red Cross Blood Transfusion Service and Laboratory for Experimental and Clinical Immunology, University of Amsterdam, Amsterdam

,

J J Abbink

The Central Laboratory of the Netherlands Red Cross Blood Transfusion Service and Laboratory for Experimental and Clinical Immunology, University of Amsterdam, Amsterdam

,

M C Brouwer

The Central Laboratory of the Netherlands Red Cross Blood Transfusion Service and Laboratory for Experimental and Clinical Immunology, University of Amsterdam, Amsterdam

,

C Meijer

The Central Laboratory of the Netherlands Red Cross Blood Transfusion Service and Laboratory for Experimental and Clinical Immunology, University of Amsterdam, Amsterdam

,

D Roem

The Central Laboratory of the Netherlands Red Cross Blood Transfusion Service and Laboratory for Experimental and Clinical Immunology, University of Amsterdam, Amsterdam

,

G P Voorn

¹The Department of Infectious Disease, University of Leiden, Delft, The Netherlands

,

J W J Lambers

²The Department of Cell, Gist Brocades, Delft, The Netherlands

,

J A van Mourik

The Central Laboratory of the Netherlands Red Cross Blood Transfusion Service and Laboratory for Experimental and Clinical Immunology, University of Amsterdam, Amsterdam

,

C E Hack

The Central Laboratory of the Netherlands Red Cross Blood Transfusion Service and Laboratory for Experimental and Clinical Immunology, University of Amsterdam, Amsterdam

› Author Affiliations

Abstract

PDF Download

Summary

The acute phase behaviour of the fast inhibitor of tissue-type plasminogen activator (PAI-1) in vivo has been attributed to increased synthesis by endothelial cells. However, most other acute phase proteins in vivo are synthesized in the liver, which process is regulated by cytokines and can be studied in the hepatoma derived cell line HepG2.

In this study, we investigated whether the synthesis of PAI-1 by HepG2 cells is regulated by the cytokines recombinant IL-1, rlL-6 and rTNF. Recombinant IL-1 and rTNF each increased PAI-1 synthesis by Hep G2 cells two to three fold, whereas rIL-6 hardly had an effect. Mixtures of rIL-1, rIL-6 and rTNF increased PAI-1 synthesis up to eleven fold. The effects observed were not due to non-specific effects on Hep G2 cell metabolism, since synthesis of alpha-2-antiplasmin was not effected by any of those cytokines, whereas fibrinogen synthesis was increased three to four fold by rIL-6, but was unaffected by rIL-1. Thus, our results demonstrate that synthesis of PAI-1 by Hep G2 cells is regulated by cytokines and implicate that the acute phase behaviour of PAI-1 in vivo at least in part may be due to an increased synthesis by the liver.

PDF (1372 kb)

References

REFERENCES
1 Kruithof EKO. Plasminogen Activator Inhibitor type 1: biochemical, biological and clinical aspects (Review). Fibrinolysis 1988; 2: 59-70
2 Sprengers ED, Kluft C. Plasminogen activator inhibitors. Blood 1987; 69: 381-387
3 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
4 van Mourik JA, Lawrence DA, Loskutoff DJ. Purification of an inhibitor of plasminogen activator (antiactivator) synthesized by endothelial cells. J Biol Chem 1984; 259: 14914-14921
5 Sprengers ED, Princen HMG, Kooistra T, Van Hinsbergh VWM. Inhibition of plasminogen activators by conditioned medium of human hepatocytes and hepatoma cell line HepG2. J Lab Clin Med 1985; 105: 751-758
6 Alessi MC, Juhan-Vague I, Kooistra T, Declerck PJ, Collen D. Insulin stimulates the synthesis of plasminogen activator inhitibor 1 by the human hepatocellular cell line Hep G2. Thromb Haemostas 1988; 60: 491-494
7 Erickson LA, Ginsberg MH, Loskutoff DJ. Detection and partial characterization of an inhibitor of plasminogen activator in human platelets. J Clin Invest 1984; 74: 1465-1472
8 Booth NA, Simpson AJ, Croll A, Bennett B, MacGregor IR. Plasminogen activator inhibitor (PAI-1) in plasma and platelets. Br J Haematol 1988; 70: 327-333
9 Lambers JWJ, Cammenga M, Konig BW, Mertens K, Pannekoek H, van Mourik JA. Activation of human endothelial cell-type plasminogen activator inhibitor (PAI-1) by negatively charged phospholipids. J Biol Chem 1987; 262: 17492-17496
10 Katagiri K, Okada K, Hattori H, Yano M. Bovine endothelial cell plasminogen activator inhibitor. Eur J Biochem 1988; 176: 81-87
11 Hekman CM, Loskutoff DJ. Endothelial cells produce a latent inhibitor of plasminogen activators that can be activated by denatur-ants. J Biol Chem 1989; 260: 11581-11587
12 Lawrence DA, Loskutoff DJ. Inactivation of plasminogen activator inhibitor by oxidants. Biochemistry 1986; 25: 6351-6355
13 Suffredini AF, Harpel PC, Parillo JE. Promotion and subsequent inhibition of plasminogen activator after administration of intravenous endotoxin to normal subjects. N Engl J Med 1989; 18: 1165-1171
14 Juhan-Vague I, Aillaud MF, de Cock F, Philip-Joet C, Arnaud C, Serradimigni A, Collen D. The fast-acting inhibitor of tissue plasminogen activator is an acute phase reactant protein. In: Progress in Fibrinolysis Davidson JF, Donati MB, Coccheri S. (eds) Churchill-Livingstone; Edinburgh: 1985: 146
15 Kluft C, Verheijen JH, Jie AFH, Rijken DC, Preston FE, Sue-Ling HM, Jespersen J, Aasen AO. The postoperative fibrinolytic shutdown: a rapidly reverting acute phase pattern for the fast-acting inhibitor of tissuc-type plasminogen activator after trauma. Scand J Clin Lab Invest 1985; 45: 605-610
16 Medina R, Socher SH, Han JH, Friedman PA. Interleukin-1, endotoxin or tumor necrosis factor/cachectin enhance the level of plasminogen activator inhibitor messenger RNA in bovine aortic endothelial cells. Thromb Res 1989; 54: 41-52
17 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
18 Nachman RL, Hajjar KA, Silverstein RL, Dinarello CA. Interleukin 1 induces endothelial cell synthesis of plasminogen activator inhibitor. J Exp Med 1986; 163: 1595-1600
19 Van Hinsbergh VWM, Kooistra T, van den Berg EA, Princen HMG, Fiers W, Emeis JJ. Tumor necrosis factor increases the production of plasminogen activator inhibitor in human endothelial cells in vitro and rats in vivo. Blood 1988; 72: 1467-1473
20 Kushner I, Mackiewicz A. Acute phase proteins as disease markers. Disease Markers 1987; 5: 1-11
21 Dowton SB, Colten HR. Acute phase reactants in inflammation and infection. Semin Hematol 1988; 2: 84-90
22 Le J, Vilcek J. Interleukin 6: A multifunctional cytokine regulating immune reactions and the acute phase protein response. Lab Invest 1989; 61: 588-602
23 Heinrich PC, Castell JV, Andus T. Interleukin-6 and the acute phase response. Biochem J 1990; 265: 621-636
24 Brakenhoff JPJ, De Groot ER, Evers RF, Pannekoek H, Aarden LA. Molecular cloning and expression of hybridoma growth factor in E. coli. J Immunol 1987; 139: 4116-4121
25 Helle M, Brakenhoff JPJ, De Groot ER, Aarden LA. Interleukin-6 is involved in Interleukin-1-induced activities. Eur J Immunol 1988; 18: 957-959
26 Helle M, Boeije L, Aarden LA. Functional discrimination between interleukin 6 and interleukin 1. Eur J Immunol 1988; 18: 1535-1540
27 Espevik T, Nissen-Meyer J. A highly sensitive cell line, WEHI 164 clone 13, for measuring cytotoxic factor/tumor necrosis from human monocytes. J Immunol Methods 1986; 95: 99-105
28 Lambers JWJ, Voorn GP, Klein GebbinkR, Pannekoek H, van Mourik JA. Monoclonal antibodies against human endothelial plasminogen activator inhibitor (PAI-1) that distinguish between the active and latent form and can be used to purify endothelial and E. coli recombinant PAI-1. Fibrinolysis 1988; 2 (Suppl. 01) 33 (Abstract)
29 Hack CE, Hannema AJ, Eerenberg AJM, Out TA, Aalberse RC. A Cl-inhibitor-complex assay (INCA): a method to detect Cl activation in vitro and in vivo. J Immunol 1981; 127: 1450-1453
30 Hunter WM, Greenwood FC. Preparation of 131-Iodine labeled human growth hormone of high specific activity. Nature 1962; 194: 495-496
31 Bachmann F. Fibrinolysis. In: Thrombosis and Haemostasis XIth Congress Verstraete M, Vermylen J, Lijnen R, Amout J. (eds) Leuven University Press; Leuven: 1987: 227
32 Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227: 680-685
33 Towbin H, Stahelin T, Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 1979; 76: 4350-4354
34 Knowles BB, Howe CC, Aden ADP. Human hepatocellular carcinoma cell lines secrete the major plasma proteins and hepatitis b surface antigen. Science 1980; 209: 497-499
35 Kushner I. Regulation of the acute phase liver gene expression. In: Regulation of the Acute Phase and Immune Responses: Interleukin 6 Seghal PB, Greininger G, Tosato G. (eds) NY Acad Sci; New York: 1989: 240
36 Sawdey M, Podor TJ, Loskutoff DJ. Regulation of type 1 plasminogen activator inhibitor gene expression in cultured bovine aortic endothelial cells. Induction by transforming growth factor-p, lipopolysaccharide, and tumor necrosis factor-a. J Biol Chem 1989; 264: 10396-10401