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J Cell Biol. 1991 Oct 1; 115(1): 191–199.
PMCID: PMC2289921

Metastatic behavior of human melanoma cell lines in nude mice correlates with urokinase-type plasminogen activator, its type-1 inhibitor, and urokinase-mediated matrix degradation


Five out of six human melanoma cell lines tested were able to degrade in vitro a smooth muscle cell extracellular matrix in a plasmin- dependent way. In three of these five cell lines, this process was mediated by tissue-type plasminogen activator (t-PA) and in the other two cell lines by urokinase-type plasminogen activator (u-PA). All melanoma cell lines produced t-PA mRNA and protein, whereas only the two cell lines showing u-PA-mediated matrix degradation produced u-PA mRNA and protein. These latter cell lines also produced plasminogen activator inhibitor type-1 (PAI-1) and type-2 (PAI-2) mRNA and protein. u-PA receptor (u-PA-R) mRNA and binding of radiolabeled u-PA was found in all melanoma cell lines. The metastatic capacity of these cell lines was studied in nude mice. All cell lines were able to develop primary tumors at the subcutaneous inoculation site. The production of plasminogen activators, their inhibitors and urokinase receptor by subcutaneous tumors corresponded with the production by the parental cell lines in vitro. The two u-PA and PAI-1 producing cell lines showed the highest frequency to form spontaneous lung metastases after subcutaneous inoculation, whereas five of the six cell lines formed lung colonies after intravenous inoculation. In conclusion, u-PA mediated matrix degradation in vitro and production of u-PA and PAI-1 by human melanoma cell lines correlated with their ability to form spontaneous lung metastasis in nude mice. No correlation was found with the ability to form lung colonies after intravenous injection. These findings suggest a role for u-PA and PAI-1 in a relatively early stage of melanoma metastasis.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Andreasen PA, Nielsen LS, Kristensen P, Grøndahl-Hansen J, Skriver L, Danø K. Plasminogen activator inhibitor from human fibrosarcoma cells binds urokinase-type plasminogen activator, but not its proenzyme. J Biol Chem. 1986 Jun 15;261(17):7644–7651. [PubMed]
  • Appella E, Robinson EA, Ullrich SJ, Stoppelli MP, Corti A, Cassani G, Blasi F. The receptor-binding sequence of urokinase. A biological function for the growth-factor module of proteases. J Biol Chem. 1987 Apr 5;262(10):4437–4440. [PubMed]
  • Axelrod JH, Reich R, Miskin R. Expression of human recombinant plasminogen activators enhances invasion and experimental metastasis of H-ras-transformed NIH 3T3 cells. Mol Cell Biol. 1989 May;9(5):2133–2141. [PMC free article] [PubMed]
  • Baker MS, Bleakley P, Woodrow GC, Doe WF. Inhibition of cancer cell urokinase plasminogen activator by its specific inhibitor PAI-2 and subsequent effects on extracellular matrix degradation. Cancer Res. 1990 Aug 1;50(15):4676–4684. [PubMed]
  • Behrendt N, Rønne E, Ploug M, Petri T, Løber D, Nielsen LS, Schleuning WD, Blasi F, Appella E, Danø K. The human receptor for urokinase plasminogen activator. NH2-terminal amino acid sequence and glycosylation variants. J Biol Chem. 1990 Apr 15;265(11):6453–6460. [PubMed]
  • Binnema DJ, van Iersel JJ, Dooijewaard G. Quantitation of urokinase antigen in plasma and culture media by use of an ELISA. Thromb Res. 1986 Sep 1;43(5):569–577. [PubMed]
  • Bizik J, Lizonová A, Stephens RW, Grófová M, Vaheri A. Plasminogen activation by t-PA on the surface of human melanoma cells in the presence of alpha 2-macroglobulin secretion. Cell Regul. 1990 Nov;1(12):895–905. [PMC free article] [PubMed]
  • Blasi F, Vassalli JD, Danø K. Urokinase-type plasminogen activator: proenzyme, receptor, and inhibitors. J Cell Biol. 1987 Apr;104(4):801–804. [PMC free article] [PubMed]
  • Cajot JF, Schleuning WD, Medcalf RL, Bamat J, Testuz J, Liebermann L, Sordat B. Mouse L cells expressing human prourokinase-type plasminogen activator: effects on extracellular matrix degradation and invasion. J Cell Biol. 1989 Aug;109(2):915–925. [PMC free article] [PubMed]
  • Chomczynski P, Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. [PubMed]
  • Cubellis MV, Nolli ML, Cassani G, Blasi F. Binding of single-chain prourokinase to the urokinase receptor of human U937 cells. J Biol Chem. 1986 Dec 5;261(34):15819–15822. [PubMed]
  • Cubellis MV, Andreasen P, Ragno P, Mayer M, Danø K, Blasi F. Accessibility of receptor-bound urokinase to type-1 plasminogen activator inhibitor. Proc Natl Acad Sci U S A. 1989 Jul;86(13):4828–4832. [PMC free article] [PubMed]
  • Cubellis MV, Wun TC, Blasi F. Receptor-mediated internalization and degradation of urokinase is caused by its specific inhibitor PAI-1. EMBO J. 1990 Apr;9(4):1079–1085. [PMC free article] [PubMed]
  • Danø K, Andreasen PA, Grøndahl-Hansen J, Kristensen P, Nielsen LS, Skriver L. Plasminogen activators, tissue degradation, and cancer. Adv Cancer Res. 1985;44:139–266. [PubMed]
  • Dodemont HJ, Soriano P, Quax WJ, Ramaekers F, Lenstra JA, Groenen MA, Bernardi G, Bloemendal H. The genes coding for the cytoskeletal proteins actin and vimentin in warm-blooded vertebrates. EMBO J. 1982;1(2):167–171. [PMC free article] [PubMed]
  • Ellis V, Scully MF, Kakkar VV. Plasminogen activation initiated by single-chain urokinase-type plasminogen activator. Potentiation by U937 monocytes. J Biol Chem. 1989 Feb 5;264(4):2185–2188. [PubMed]
  • Ellis V, Wun TC, Behrendt N, Rønne E, Danø K. Inhibition of receptor-bound urokinase by plasminogen-activator inhibitors. J Biol Chem. 1990 Jun 15;265(17):9904–9908. [PubMed]
  • Estreicher A, Wohlwend A, Belin D, Schleuning WD, Vassalli JD. Characterization of the cellular binding site for the urokinase-type plasminogen activator. J Biol Chem. 1989 Jan 15;264(2):1180–1189. [PubMed]
  • Gaffney PJ, Curtis AD. A collaborative study of a proposed international standard for tissue plasminogen activator (t-PA). Thromb Haemost. 1985 Feb 18;53(1):134–136. [PubMed]
  • Gavrilovic J, Murphy G. The role of plasminogen in cell-mediated collagen degradation. Cell Biol Int Rep. 1989 Apr;13(4):367–375. [PubMed]
  • Goldfarb RH, Liotta LA. Proteolytic enzymes in cancer invasion and metastasis. Semin Thromb Hemost. 1986 Oct;12(4):294–307. [PubMed]
  • Goldfarb RH, Ziche M, Murano G, Liotta LA. Plasminogen activators (urokinase) mediate neovascularization: possible role in tumor angiogenesis. Semin Thromb Hemost. 1986 Oct;12(4):337–338. [PubMed]
  • Grøndahl-Hansen J, Ralfkiaer E, Kirkeby LT, Kristensen P, Lund LR, Danø K. Localization of urokinase-type plasminogen activator in stromal cells in adenocarcinomas of the colon in humans. Am J Pathol. 1991 Jan;138(1):111–117. [PMC free article] [PubMed]
  • Hearing VJ, Law LW, Corti A, Appella E, Blasi F. Modulation of metastatic potential by cell surface urokinase of murine melanoma cells. Cancer Res. 1988 Mar 1;48(5):1270–1278. [PubMed]
  • Hébert CA, Baker JB. Linkage of extracellular plasminogen activator to the fibroblast cytoskeleton: colocalization of cell surface urokinase with vinculin. J Cell Biol. 1988 Apr;106(4):1241–1247. [PMC free article] [PubMed]
  • Jones PA, DeClerck YA. Destruction of extracellular matrices containing glycoproteins, elastin, and collagen by metastatic human tumor cells. Cancer Res. 1980 Sep;40(9):3222–3227. [PubMed]
  • Jones PA, Scott-Burden T, Gevers W. Glycoprotein, elastin, and collagen secretion by rat smooth muscle cells. Proc Natl Acad Sci U S A. 1979 Jan;76(1):353–357. [PMC free article] [PubMed]
  • Katano M, Saxton RE, Cochran AJ, Irie RF. Establishment of an ascitic human melanoma cell line that metastasizes to lung and liver in nude mice. J Cancer Res Clin Oncol. 1984;108(2):197–203. [PubMed]
  • Kristensen P, Larsson LI, Nielsen LS, Grøndahl-Hansen J, Andreasen PA, Danø K. Human endothelial cells contain one type of plasminogen activator. FEBS Lett. 1984 Mar 12;168(1):33–37. [PubMed]
  • Kruithof EK. Plasminogen activator inhibitors--a review. Enzyme. 1988;40(2-3):113–121. [PubMed]
  • Liotta LA. Tumor invasion and metastases--role of the extracellular matrix: Rhoads Memorial Award lecture. Cancer Res. 1986 Jan;46(1):1–7. [PubMed]
  • Lockshin A, Giovanella BC, De Ipolyi PD, Williams LJ, Jr, Mendoza JT, Yim SO, Stehlin JS., Jr Exceptional lethality for nude mice of cells derived from a primary human melanoma. Cancer Res. 1985 Jan;45(1):345–350. [PubMed]
  • Lund LR, Riccio A, Andreasen PA, Nielsen LS, Kristensen P, Laiho M, Saksela O, Blasi F, Danø K. Transforming growth factor-beta 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 May;6(5):1281–1286. [PMC free article] [PubMed]
  • Medcalf RL, Van den Berg E, Schleuning WD. Glucocorticoid-modulated gene expression of tissue- and urinary-type plasminogen activator and plasminogen activator inhibitor 1 and 2. J Cell Biol. 1988 Mar;106(3):971–978. [PMC free article] [PubMed]
  • Moscatelli D, Rifkin DB. Membrane and matrix localization of proteinases: a common theme in tumor cell invasion and angiogenesis. Biochim Biophys Acta. 1988 Aug 3;948(1):67–85. [PubMed]
  • Murphy G, Reynolds JJ, Hembry RM. Metalloproteinases and cancer invasion and metastasis. Int J Cancer. 1989 Oct 15;44(4):757–760. [PubMed]
  • Nakane PK, Pierce GB., Jr Enzyme-labeled antibodies: preparation and application for the localization of antigens. J Histochem Cytochem. 1966 Dec;14(12):929–931. [PubMed]
  • Nielsen LS, Kellerman GM, Behrendt N, Picone R, Danø K, Blasi F. A 55,000-60,000 Mr receptor protein for urokinase-type plasminogen activator. Identification in human tumor cell lines and partial purification. J Biol Chem. 1988 Feb 15;263(5):2358–2363. [PubMed]
  • Ossowski L. In vivo invasion of modified chorioallantoic membrane by tumor cells: the role of cell surface-bound urokinase. J Cell Biol. 1988 Dec;107(6 Pt 1):2437–2445. [PMC free article] [PubMed]
  • Ossowski L, Reich E. Antibodies to plasminogen activator inhibit human tumor metastasis. Cell. 1983 Dec;35(3 Pt 2):611–619. [PubMed]
  • 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 Dec;103(6 Pt 1):2411–2420. [PMC free article] [PubMed]
  • Pöllänen J, Saksela O, Salonen EM, Andreasen P, Nielsen L, Danø K, Vaheri A. Distinct localizations of urokinase-type plasminogen activator and its type 1 inhibitor under cultured human fibroblasts and sarcoma cells. J Cell Biol. 1987 Apr;104(4):1085–1096. [PMC free article] [PubMed]
  • Pöllänen J, Hedman K, Nielsen LS, Danø K, Vaheri A. Ultrastructural localization of plasma membrane-associated urokinase-type plasminogen activator at focal contacts. J Cell Biol. 1988 Jan;106(1):87–95. [PMC free article] [PubMed]
  • Pöllänen J, Vaheri A, Tapiovaara H, Riley E, Bertram K, Woodrow G, Stephens RW. Prourokinase activation on the surface of human rhabdomyosarcoma cells: localization and inactivation of newly formed urokinase-type plasminogen activator by recombinant class 2 plasminogen activator inhibitor. Proc Natl Acad Sci U S A. 1990 Mar;87(6):2230–2234. [PMC free article] [PubMed]
  • Quax PH, van Leeuwen RT, Verspaget HW, Verheijen JH. Protein and messenger RNA levels of plasminogen activators and inhibitors analyzed in 22 human tumor cell lines. Cancer Res. 1990 Mar 1;50(5):1488–1494. [PubMed]
  • Rifkin DB, Tsuboi R, Mignatti P. The role of proteases in matrix breakdown during cellular invasion. Am Rev Respir Dis. 1989 Oct;140(4):1112–1113. [PubMed]
  • Rijken DC, Wijngaards G, Welbergen J. Immunological characterization of plasminogen activator activities in human tissues and body fluids. J Lab Clin Med. 1981 Apr;97(4):477–486. [PubMed]
  • 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 Feb;9(2):467–474. [PMC free article] [PubMed]
  • Schleuning WD, Medcalf RL, Hession C, Rothenbühler R, Shaw A, Kruithof EK. Plasminogen activator inhibitor 2: regulation of gene transcription during phorbol ester-mediated differentiation of U-937 human histiocytic lymphoma cells. Mol Cell Biol. 1987 Dec;7(12):4564–4567. [PMC free article] [PubMed]
  • Sprengers ED, Kluft C. Plasminogen activator inhibitors. Blood. 1987 Feb;69(2):381–387. [PubMed]
  • Stephens RW, Pöllänen J, Tapiovaara H, Leung KC, Sim PS, Salonen EM, Rønne E, Behrendt N, Danø K, Vaheri A. Activation of pro-urokinase and plasminogen on human sarcoma cells: a proteolytic system with surface-bound reactants. J Cell Biol. 1989 May;108(5):1987–1995. [PMC free article] [PubMed]
  • Stoppelli 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 U S A. 1985 Aug;82(15):4939–4943. [PMC free article] [PubMed]
  • Stoppelli MP, Tacchetti C, Cubellis MV, Corti A, Hearing VJ, Cassani G, Appella E, Blasi F. Autocrine saturation of pro-urokinase receptors on human A431 cells. Cell. 1986 Jun 6;45(5):675–684. [PubMed]
  • Tryggvason K, Höyhtyä M, Salo T. Proteolytic degradation of extracellular matrix in tumor invasion. Biochim Biophys Acta. 1987 Nov 25;907(3):191–217. [PubMed]
  • van den Berg EA, Sprengers ED, Jaye M, Burgess W, Maciag T, van Hinsbergh VW. Regulation of plasminogen activator inhibitor-1 mRNA in human endothelial cells. Thromb Haemost. 1988 Aug 30;60(1):63–67. [PubMed]
  • van Muijen GN, Cornelissen IM, Jansen CF, Ruiter DJ. Progression markers in metastasizing human melanoma cells xenografted to nude mice (review). Anticancer Res. 1989 Jul-Aug;9(4):879–884. [PubMed]
  • van Muijen GN, Jansen KF, Cornelissen IM, Smeets DF, Beck JL, Ruiter DJ. Establishment and characterization of a human melanoma cell line (MV3) which is highly metastatic in nude mice. Int J Cancer. 1991 Apr 22;48(1):85–91. [PubMed]
  • van Zonneveld AJ, Chang GT, van den Berg J, Kooistra T, Verheijen JH, Pannekoek H, Kluft C. Quantification of tissue-type plasminogen activator (t-PA) mRNA in human endothelial-cell cultures by hybridization with a t-PA cDNA probe. Biochem J. 1986 Apr 15;235(2):385–390. [PMC free article] [PubMed]
  • Vassalli JD, Baccino D, Belin D. A cellular binding site for the Mr 55,000 form of the human plasminogen activator, urokinase. J Cell Biol. 1985 Jan;100(1):86–92. [PMC free article] [PubMed]
  • Verheijen JH, Mullaart E, Chang GT, Kluft C, Wijngaards G. A simple, sensitive spectrophotometric assay for extrinsic (tissue-type) plasminogen activator applicable to measurements in plasma. Thromb Haemost. 1982 Dec 27;48(3):266–269. [PubMed]
  • Versteeg R, Noordermeer IA, Krüse-Wolters M, Ruiter DJ, Schrier PI. c-myc down-regulates class I HLA expression in human melanomas. EMBO J. 1988 Apr;7(4):1023–1029. [PMC free article] [PubMed]

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