Distinct Patterns of Urokinase Receptor (uPAR) Expression by Leukemic Cells and Peripheral Blood Cells

Mercè Jardi; Julia Inglés-Esteve; Maria Burgal; Carmen Azqueta; Francisco Velasco; Chary López-Pedrera; Lindsey Miles; Jordi Félez

doi:10.1055/s-0038-1650701

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 1996; 76(06): 1009-1019
DOI: 10.1055/s-0038-1650701

Original Article

Schattauer GmbH Stuttgart

Distinct Patterns of Urokinase Receptor (uPAR) Expression by Leukemic Cells and Peripheral Blood Cells

Mercè Jardi

¹Institut Recerca Oncologica Hospital Duran i Reynals, Barcelona, Spain

,

Julia Inglés-Esteve

¹Institut Recerca Oncologica Hospital Duran i Reynals, Barcelona, Spain

,

Maria Burgal

²Instituto de Investigaciones Citologicas, Valencia, Spain

,

Carmen Azqueta

¹Institut Recerca Oncologica Hospital Duran i Reynals, Barcelona, Spain

,

Francisco Velasco

³Servicio de Hematologla, Hospital Reina Sofia, Córdoba, Spain

,

Chary López-Pedrera

³Servicio de Hematologla, Hospital Reina Sofia, Córdoba, Spain

,

Lindsey Miles

⁴Department of Vascular Biology, The Scripps Research Institute, La Jolla, CA, USA

,

Jordi Félez

¹Institut Recerca Oncologica Hospital Duran i Reynals, Barcelona, Spain

› Author Affiliations

Abstract

PDF Download

Summary

The urinary type plasminogen activator, urokinase (uPA) is localized on the cell surface through the binding of a specific receptor, the uPA receptor (uPAR). The uPA localization enhances plasmin formation on the cell surface and facilitates cell migration. The cellular and tissue distribution of uPAR is not fully established. We have analyzed uPAR expression in nine leukemic cell lines of distinct lineages and maturational states and correlated this with expression of plasminogen receptors, tissue-type plasminogen activator (tPA) receptors and LDL receptor-related protein (LRP). The most immature and least differentiated cell line (an erythro-myeloid cell line) and cells of lymphoid lineage, did not express uPAR, whereas cells differentiated along the myelo-monocytic pathway displayed this receptor. Plasminogen and tPA receptors were expressed by all leukemic cell lines and by all nucleated peripheral blood cells but B and T lymphocytes were negative for cell surface expression of both uPAR and LRP while monocytes and neutrophils were positive for expression of both uPAR and LRP. PMA stimulation induced surface expression of uPAR in lymphocytes but did not induce expression of LRP by these cells. In contrast, lymphoid cell lines were negative for uPAR expression even after PMA stimulation, indicating differences in regulation of uPAR expression between lymphocytes and lymphoid cell lines. The pattern of uPAR expression on leukemic cell lines was also studied on bone marrow blast cells from leukemic patients. Only the most mature myeloid cells expressed uPAR on their surfaces. In contrast, M3 leukemic cells and other blast cells displaying lymphoid markers such as TdT (+) and/or CD2 (+) did not express intracellular or cell-surface associated uPAR, indicating an heterogeneity among these promyelo-cytic cells and suggesting that uPAR may be a useful marker for leukemia typing. Myeloid blast cells from some patients contained intracellular pools of uPAR but displayed no receptor on the cell surface, suggesting that translocation may be a mechanism regulating uPAR expression in these cells. The comparison of uPAR expression between these cell lines and peripheral blood cells and it correlation with plasminogen receptors, tPA receptors and LRP expression offers new insights regarding potential mechanisms for regulation of uPA-uP-AR-mediated pericellular proteolysis.

PDF (573 kb)

References

References
1 Nielsen LS, Kellerman GM, Behrendt N, Picone R, Dannϕ K, Blasi F. A 55,000-60,000 Mr receptor protein for urokinase-type plasminogen activator. J Biol Chem 1988; 263: 2358-2363
2 Behrendt N, Ronne E, Ploug M, Petri T, Lober D, Nielsen LS, Schleuning W-D, Blasi F, Appella E, Danϕ K. The human receptor for urokinase plasminogen activator. NH₂-terminal amino acid sequence and glycosyla-tion variants. J Biol Chem 1990; 265: 6453-6460
3 Ploug M, Ronne E, Behrendt N, Jensen AL, Blasi F, Dannϕ K. Cellular receptor for urokinase plasminogen activator. Carboxyl-terminal processing and membrane anchoring by glycosy 1-phosphatidylinositol. J Biol Chem 1991; 266: 1926-1933
4 Blasi F. Urokinase and urokinase receptor: a paracrine/autocrine system regulating cell migration and invasiveness. Bio Essays 1993; 15: 105-111
5 Pollanen J. The N-Terminal domain of human urokinase receptor contains two distinct regions critical for ligand recognition. Blood 1993; 82: 2719
6 Estreicher A, Wohlwend A, Belin D, Scleuning W, Vassalli JD. Characterization of the cellular binding site for the urokinase-type plasminogen activator. J Biol Chem 1989; 264: 1180-1189
7 Plow EF, Felez J, Miles LA. Cellular Regulation of Fibrinolysis. Thromb Haemost 1991; 66: 32-36
8 Vassalli J. The urokinase receptor. Fibrinolysis 1994; 8: 172-181
9 Blasi F, Conese M, Moller LB, Pedersen N, Cavallaro U, Cubellis MV, Fazioli F, Hemandez-Marrero L, Limongi P, Munoz-Canoves P, Resnati M, Riitinen L, Sidenius N, Soravia E, Soria MR, Stoppelli MP, Talarico D, Teesalu T, Valcamonica S. The Urokinase Receptor: Structure, Regulation and Inhibitor-mediated Internalization. Fibrinolysis 1994; 8: 182-188
10 Danø K, Behrendt N, Brunner N, Ellis V, Ploug M, Pyke C. The urokinase receptor: Protein structure and role in plasminogen activation and cancer invasion. Fibrinolysis 1994; 8: 189-203
11 Pedersen H, Brunner N, Francis D, Osterlind K, Ronne E, Hansen HH, Danø K, Grondahl-Hansen J. Prognostic impact of urokinase, urokinase receptor, and type 1 plasminogen activator inhibitor in squamous and large cell lung cancer tissue. Cancer Res 1994; 54: 4671-4675
12 Carriero MV, Franco P, Del Vecchio S, Massa O, Botti G, D,Aiuto G, Stoppelli MP, Salvatore M. Tissue distribution of soluble and receptor-bound urokinase in human breast cancer using a panel of monoclonal antibodies. Cancer Res 1994; 54: 5445-5454
13 Mazzieri R, Hoyer-Hansen G, Ronne E, Lober D, Vagnarelli P, Raimondi E, De Carli L, Danø K, Mignatti P. Urokinase and urokinase receptor expression in somatic cell hybrids. Fibrinolysis 1994; 8: 344-352
14 Mohanam S, Sawaya RE, Yamamoto M, Bruner JM, Nicholson GL, Rao JS. Proteolysis and invasiveness of brain tumors: Role of urokinase-type plasminogen activator receptor. J Neurooncol 1994; 22: 153-160
15 Semi U, Fibbi G, Anichini E, Zamperini A, Pucci M, Mannoni A, Matucci A, Benucci M, Del Rosso A, Del Rosso M. Plasminogen activator and receptor in osteoarthritis. J Rheumatol 1995; 22 43: 120-122
16 Pyke C, Ralfkiaer E, Ronne E, Hoyer-Hansen G, Kirkeby L, Dano K. Immunohistochemical detection of the receptor for urokinase plasminogen activator in human colon cancer. Histopathology 1994; 24: 131-138
17 Bianchi E, Cohen RL, Thor AT, Todd RFIII, Mizukami IF, Lawrence DA, Ljung BM, Shuman MA, Smith HS. The urokinase receptor is expressed in invasive breast cancer but not in normal breast tissue. Cancer Res 1994; 54: 861-866
18 Ronne E, Hoyer-Hansen G, Brünner N, Pedersen H, Rank F, Osborne CK, Clark GM, Danø K, Grondahl-Hansen J. Urokinase receptor in breast cancer tissue extracts. Enzyme-linked immunosorbent assay with a combination of mono- and polyclonal antibodies. Breast Cancer Res Treat 1995; 33: 199-207
19 Nykjaer A, Moller B, Todd RFIII, Christensen T, Andreasen PA, Glie-mann J, Petersen CM. Urokinase receptor: An activation antigen in human T lymphocytes. J Immunol 1994 152: 505-516
20 Plesner T, Ralfkiaer E, Wittrup M, Johnsen H, Pyke C, Pedersen TL, Hansen NE, Danp K. Expression of the receptor for urokinase-type plasminogen activator in normal and neoplastic blood cells and hematopoietic tissue. Am J Clin Pathol 1994; 102: 835-841
21 Nykjaer A, Petersen CM, Moller B, Andreasen PA, Gliemann J. Identification and characterization of urokinase receptors in natural killer cells and T-cell-derived lymphokine activated killer cells. FEBS Lett 1992; 300: 13-17
22 Nykjaer A, Petersen CM, Christensen EI, Davidsen O, Gliemann J. Urokinase receptors in human monocytes. Biochim Biophys Acta 1990; 1052: 399-407
23 Miles LA, Plow EF. Receptor mediated binding of the fibrinolytic components, plasminogen and urokinase, to peripheral blood cells. Thromb Haemost 1987; 58: 936-942
24 Todd RFIII, Bamathan ES, Bohuslav J, Chapman HA, Cohen RL, Felez J, Howell A, Johnson JG, Knapp W, Kramer M, Miles LA, Nykjaer A, Ralf-kiaer E, Shuren E. M15 Cluster. In: Leucocyte Typing V: White Cell Differentiation Antigens Schloosman SF, Boumsell L, Gilks W, Harlan JM, Kishimoto T, Morimoto C, Ritz J, Shaw S, Silverstein RL, Springer TA, Tedder TF, Todd RFIII. eds Oxford; Oxford University Press: 1995: 932-939
25 Jardi M, Ingles-Esteve J, Fabregas P, Felez J. Different expression of urokinase receptors (uPAR) among leukemic cells. Thromb Haemost 1993; 69: 608-A:231 Abstract
26 Plesner T, Ploug M, Ellis V, Ronne E, Hoyer-Hansen G, Wittrup M, Lind-hardt Pedersen T, Tscheming T, Danø K, Hansen NE. The receptor for urokinase-type plasminogen activator and urokinase is translocated from two distinct intracellular compartments to the plasma membrane on stimulation of human neutrophils. Blood 1994; 83: 808-815
27 Deutsch DG, Mertz ET. Plasminogen: Purification from human plasma by affinity chromatography. Science 1970; 170: 1995-1996
28 Miles LA, Plow EF. Topography of the high-affinity lysine binding site of plasminogen as defined with a specific antibody probe. Biochemistry 1986; 25: 6926-6933
29 Schlossman SF, Boumsell L, Gilks W, Harlan JM, Kishimoto T, Morimoto C, Ritz J, Shaw S, Silverstein RL, Springer TA, Tedder TF, Todd RF. CD antigens 1993. J Immunol 1994; 152: 1
30 Schlossman SF, Boumsell L, Gilks W, Harlan JM, Kishimoto T, Morimoto C, Ritz J, Shaw S, Silverstein RL, Springer TA, Tedder TF, Todd RF. CD antigens 1993. Blood 1994; 83: 879-880
31 Plow EF, Freaney DE, Plescia J, Miles LA. The plasminogen system and cell surfaces: Evidence for plasminogen and urokinase receptors on the same cell type. J Cell Biol 1986; 103: 2411-2420
32 Phillips DR, Morrison M. Exposed protein on the intact human erythrocyte. Biochemistry 1971; 10: 1766-1771
33 Félez J, Chanquia CJ, Fabregas P, Plow EF, Miles LA. Competition between Plasminogen and t-PA for Cellular Binding Sites. Blood 1993; 82: 2433-2441
34 Félez J, Chanquia CJ, Levin EG, Miles LA, Plow EF. Binding of tissue plasminogen activator to human monocytes and monocytoid cells. Blood 1991; 78: 2318-2327
35 Félez J, Miles LA, Plescia J, Plow EF. Regulation of Plasminogen Receptor Expression on Human Monocytes and Monocytoid Cell Lines. J Cell Biol 1990; 111: 1673-1683
36 Miles LA, Plow EF. Binding and activation of plasminogen on the platelet surface. J Biol Chem 1985; 260: 4303-4311
37 Miles LA, Ginsberg MH, White JG, Plow EF. Plasminogen interacts with human platelets through two distinct mechanisms. J Clin Invest 1986; 77: 2001-2009
38 Bennett JM, Catovsky D, Daniel MT. Proposed revised criteria for the classification of acute myeloid leukemia: A report of the French-American-British Cooperative Group. Ann Intern Med 1985; 103: 626-629
39 Roldan AL, Cubellis MV, Masucci MT, Behrendt N, Lund LR, Danø K, Appella E, Blasi F. Cloning and expression of the receptor for human urokinase plasminogen activator, a central molecule in cell surface, plasmin dependent proteolysis. EMBO J 1990; 9: 467-474
40 Lund LR, Ronne E, Roldan AL, Behrendt N, Romer J, Blasi F, Danø K. Urokinase receptor mRNA level and gene transcription are strongly and rapidly increased by phorbol myristate acetate in human monocyte-like U937 cells. J Biol Chem 1991; 266: 5177-5181
41 McCrae KR, Zini JM, Mazar A, Henkin J, Graham C, Lala PK. Human placental trophoblasts express receptors for single chain urokinase-type plasminogen activator. Third Inter.Workshop Molec & Cellul Biol of Plasminogen Activation 1991 1. 9 Abstract
42 Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 1976; 72: 248-254
43 Hoyer-Hansen G, Ronne E, Solberg H, Behrendt N, Ploug M, Lund LR, Ellis V, Danø K. Urokinase plasminogen activator cleaves its cell surface receptor releasing the ligand-binding domain. J Biol Chem 1992; 267: 18224-18229
44 Herz J, Clouthier DE, Hammer RE. LDL receptor-related protein internalizes and degrades uPA-PAI-1 complexes and is essential for embryo implantation. Cell 1992; 71: 411-421
45 Strickland DK, Kounnas MZ, Williams SE, Argraves WS. LDL receptor-related protein (LRP): A multiligand receptor. Fibrinolysis 1994; 8 (Suppl. 01) 204-215
46 Li H, Kuo A, Kochan J, Strickland D , Kariko K, Bamathan ES, Cines DB. Endocytosis of urokinase-plasminogen activator inhibitor type 1 complexes bound to a chimeric transmembrane urokinase receptor. J Biol Chem 1994; 269: 8153-8158
47 Nykjaer A, Petersen CM, Moller B, Jensen PA, Moestmp SK, Holtet TL, Etzerodt M, Thogersen HC, Munch M, Andreasen PA, Gliemann J. Purified alpha-2-macroglobulin receptor/LDL receptor-related protein binds urokinase-plasminogen activator inhibitor type-1 complex: evidence that the alpha-2-macroglobulin receptor mediates cellular degradation of urokinase receptor-bound complexes. J Biol Chem 1992; 267: 14543-14546
48 Lopez-Pedrera C, Velasco F, Dorado G, Torres A, Felez J. Tissue factor expression in leukemic cells correlates with myeloid differentiation. Haemostasis 1994; 24: 309
49 Picone R, Kajitaniak EL, Nielsen LS, Behrendt N, Mastronicola MR, Cubellis MV, Stoppelli MP, Pedersen S, Dan K, Blasi F. Regulation of urokinase receptors in monocytelike U937 cells by phorbol ester phorbol myristate acetate. J Cell Biol 1989; 108: 693-702
50 Kirchheimer JC. Modulation of receptor bound urokinase-type plaspiino-gen activator on human monocytes by non-steroidal antiinflammatory drugs. Scand J Rheumatol 1993; 22: 53-7
51 Lu H, Li H, Mirshahi SS, Soria C, Soria J, Menashi S. Comparative study of fibrinolytic activity on 937 line after stimulation by interferon gamma, 1,25 dihydroxy vitamin D₃ and their combination. Thromb Res 1993; 69: 353-359
52 Nishizuka Y. Studies and perspectives of protein kinase C. Science 1986; 233: 305-312
53 Stopelli MP, Corti A, Soffientini A, Cassani G, Blasi F, Assoian RK. Differentiation-enhanced binding of the amino-terminal fragment of human urokinase plasminogen activator to a specific receptor on U937 monocytes. Proc Natl Acad Sci USA 1985; 82: 4939-4943
54 Sherman LA, Lee J. Specific binding of soluble fibrin to macrophages. J Exp Med 1977; 145: 76-85
55 Alitalo R. Induced differentiation of K562 leukemia cells: A model for studies of gene expression in early megakaryoblasts. Leukemia Res 1990; 14: 501-514
56 Miles LA, Fowler BJ, Jardi M, Schleef RR, Felez J. Comparison of urokinase receptor expression in platelets and megakaryocytes. Blood 1995; 86 (Suppl. 01) 556a (Abstr)
57 Plow EF, Miles LA. Plasminogen receptors in the mediation of pericellular proteolysis. Cell Differ Dev 1990; 32: 293-298
58 Miles LA. Regulation of fibrinolysis and proteolysis by plasminogen receptors. Biol Clin Hematol 1993; 15: 107-115
59 Felez J, Miles LA, Fabregas P, Jardi M, Plow EF, Lijnen HR. Characterization of Cellular Binding Sites and Interactive Regions within Reactants Required for Enhancement of Plasminogen Activation by tPA on the Surface of Leukocytic Cells. Thromb Haemost 1996; 76: 577-584
60 Lopez-Alemany R, Longstaff C, Fabregas P, Jardi M, Merton E, Felez J. Inhibition of plasmin generation on cell surfaces by monoclonal antibodies against a 55-60 kDa plasminogen receptor. Fibrinolysis 1996; 10: 5-A-14 Abstract
61 Vassalli JD, Pepper SM. Membrane Protease in Focus. Nature 1994; 370: 14-15
62 Pepper MS, Sappino AP, Stocklin R, Montesano R, Orci L, Vassalli JD. Upregulation of urokinase receptor expression on migrating endothelial cells. J Cell Biol 1993; 122: 673-684
63 Conese M, Blasi F. Urokinase/urokinase receptor system: Internalization/degradation of urokinase-serpin complexes: Mechanism and regulation. Biol Chem Hoppe Seyler 1995; 376: 143-155
64 Claxton DF, Reading CL, Nagarajan L, Tsujimoto Y, Andersson BS, Estev E, Cork A, Huh YO, Trujillo J, Deisseroth AB. Correlation of CD2 Expression with PML Gene Breakpoints in patients with acute promyelo-cytic leukemia. Blood 1992; 80: 582-586
65 Moller LB, Pöllänen J, Ronne E, Pedersen N, Blasi F. N-linked glyco-sylation of the ligand-binding domain of the human urokinase receptor contributes to the affinity for its ligand. J Biol Chem 1993; 268: 11152-11159
66 Pyke C, Graem N, Ralfkiaer E, Ronne E, Hoyer-Hansen G, Brünner N, Danø K. Receptor for urokinase is present in tumor-associated macrophages in ductal breast carcinoma. Cancer Res 1993; 53: 1911-1915
67 Pyke C, Kristensen P, Palkfjaer E, Grondhal-Hansen J, Eriksen J, Blasi F, Danø K. Urokinase-type plasminogen activator is expressed in stromal cells and its receptor in cancer cells at invasive foci in human colon adenocarcinomas. Am J Pathol 1991; 138: 1059-1067