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. 1993 Jun;84(6):633–640. doi: 10.1111/j.1349-7006.1993.tb02023.x

Increased Cell‐surface Urokinase in Advanced Ovarian Cancer

Hiroshi Kobayashi 1, Nobuhiko Moniwa 1, Motoi Sugimura 1, Hiromitsu Shinohara 1, Hidekazu Ohi 1, Toshihiko Terao 1
PMCID: PMC5919317  PMID: 8340251

Abstract

Urokinase‐type plasminogen activator (uPA), uPA receptors, and cathepsin B were quantitatcd by using an immunological method, enzyme‐linked immunosorbent assay, and amidolytic activity assays in 15 malignant and 10 benign epithelial ovarian tumors. The levels of uPA and uPA receptors, as well as cathepsin B, were found to be higher in membrane preparations obtained from malignant tumors than in those obtained from benign tumors. Acid‐treated membranes acquired the ability to bind uPA, indicating that uPA is bound to a specific surface receptor that is not completely saturated. Levels of single‐chain uPA (pro‐uPA) and high‐molecular‐weight uPA in membrane preparations were measured by immunoadsorbent‐amidolytic assay. The finding of a significant increase in amidolytic activity following activation of uPAs by plasmin suggested that less than half (30–40%) of all membrane immunoreactive uPAs is present in the enzymatically inactive pro‐uPA form. In the membranes of malignant tumors, levels of uPA receptor and cathepsin B did not vary with stage of disease. On the other hand, we found that the level of receptor‐bound uPA antigen/activity was significantly increased in advanced malignant tumors. Receptor‐bound uPA may play an important role in determining invasive potential of tumor cells. Since ovarian cancer cells produce both pro‐uPA and cathepsin B, the possibility of activation of tumor cell‐derived pro‐uPA by cellular protease cathepsin B must be considered.

Keywords: Urokinase‐type plasminogen activator (uPA), Single‐chain uPA, uPA receptor, Cathepsin B, Ovarian cancer

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REFERENCES

  • 1. ) Dano , K. , Andreasen , P. A. , Grondahl‐Hansen , J. , Kristensen , P. I. , Nielsen , L. S. and Skriver , L.Plasminogen activators, tissue degradation, and cancer . Adv. Cancer Res. , 44 , 139 – 266 ( 1985. ). [DOI] [PubMed] [Google Scholar]
  • 2. ) Liotta , L. A. , Rao , C. N. and Barsky , S. H.Tumor invasion and the extracellular matrix . Lab. Invest. , 49 , 636 – 649 ( 1983. ). [PubMed] [Google Scholar]
  • 3. ) Appella , E. , Robinson , E. A. , Ullrich , S. J. , Stoppelli , M. P. , Corti , A. , Cassani , G. and Blasi , F.The receptorbinding sequence of unokinase . J. Biol. Chem. , 262 , 4437 – 4440 ( 1987. ). [PubMed] [Google Scholar]
  • 4. ) Stoppelli , M. P. , Tacchetti , C. , Cubellis , M. V. , Corti , A. , Hearing , V. J. , Cassani , G. , Appella , E. and Blasi , F.Autocrine saturation of pro‐urokinase receptors on human A431 cells . Cell , 45 , 675 – 684 ( 1986. ). [DOI] [PubMed] [Google Scholar]
  • 5. ) Kobayashi , H. , Schmitt , M. , Goretzki , L. , Chucholowski , N. , Calvete , J. , Kramer , M. , Gunzler , W. A. , Janicke , F. and Graeff , H.Cathepsin B efficiently activates the soluble and the tumor cell receptor‐bound form of the proenzyme urokinase‐type plasminogen activator (pro‐uPA) . J. Biol. Chem. , 266 , 5147 – 5152 ( 1991. ). [PubMed] [Google Scholar]
  • 6. ) Goretzki , L. , Schmitt , M. , Mann , K. , Calvete , V. , Chucholowski , N. , Kramer , M. , Gunzler , W. A. , Janicke , F. and Graeff , H.Effective activation of the proenzyme form of the urokinase‐type plasminogen activator (pro‐uPA) by the cysteine protease cathepsin L . FEBS Lett. , 297 , 112 – 118 ( 1992. ). [DOI] [PubMed] [Google Scholar]
  • 7. ) Sloane , B. F. , Honn , K. V. , Sadler , J. G. , Turner , W. A. , Kimpson , J. J. and Taylor , J. D.Cathepsin B activity in B16 melanoma cells: a possible marker for metastatic potential . Cancer Res. , 42 , 980 – 986 ( 1982. ). [PubMed] [Google Scholar]
  • 8. ) Rozhin , J. , Robinson , D. , Stevens , M. A. , Lah , T. T. , Honn , K. V. , Ryan , R. E. and Sloane , B. F.Properties of a plasma membrane‐associated cathepsin B‐like cysteine proteinase in metastatic B16 melenoma . Cancer Res. , 47 , 6620 – 6628 ( 1987. ). [PubMed] [Google Scholar]
  • 9. ) Keren , Z. and LeGrue , S. J.Identification of cell surface cathepsin B‐like activity on murine melanomas and fibrosarcomas: modulation by butanol extraction . Cancer Res. , 48 , 1416 – 1521 ( 1988. ). [PubMed] [Google Scholar]
  • 10. ) Eckhout , Y. and Vaes , G.Further studies on the activation of procollagenase, the latent precursor of bone collagenase: effect of lysosomal cathepsin B. plasmin, and kallikrein, and spontaneous activation . Biochem. J. , 166 , 21 – 31 ( 1977. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 11. ) Kobayashi , H. , Ohi , H. , Sugimura , M. , Shinohara , H. , Fujii , T. and Terao , T.Inhibition of in vitro ovarian cancer cell invasion by modulation of urokinase‐type plasminogen activator and cathepsin B . Cancer Res. , 52 , 3610 – 3614 ( 1992. ). [PubMed] [Google Scholar]
  • 12. ) Casslén , B. , Gustavsson , B. and Astedt , B.Cell membrane receptors for urokinase plasminogen activator are increased in malignant ovarian tumours . Eur. J. Cancer , 27 , 1445 – 1448 ( 1991. ). [DOI] [PubMed] [Google Scholar]
  • 13. ) Bradford , B. M.A rapid and sensitive method for the quantitation of microgram quantities of protein utilising the principle of protein‐dye binding . Anal. Biochem. , 72 , 248 – 254 ( 1976. ). [DOI] [PubMed] [Google Scholar]
  • 14. ) Guesdon , J. L. , Ternynck , T. and Avrameas , S.The use of avidin‐biotin interaction in immunoenzymatic techniques . J. Histochem. Cytochem. , 27 , 1131 – 1139 ( 1979. ). [DOI] [PubMed] [Google Scholar]
  • 15. ) Solberg , H. , Lober , D. , Eriksen , J. , Ploug , M. , Ronne , E. , Behrendt , N. , Dana , K. and Hoyer‐Hansen , G.Identification and characterization of the murine cell surface receptor for the urokinase‐type plasminogen activator . Ear. J. Biochem. , 205 , 451 – 458 ( 1992. ). [DOI] [PubMed] [Google Scholar]
  • 16. ) Corti , A. , Nolli , M. L. and Cassani , G.Differential detection of single‐chain and two‐chain urokinase‐type plasminogen activator by a new immunoadsorbent‐amidolytic assay (IAA) . Thromb. Haemostasia , 56 , 407 – 410 ( 1986. ). [PubMed] [Google Scholar]
  • 17. ) Weiss , R. E. , Liu , B. C.‐S. , Ahlering , T. , Dubeau , L. and Droller , M. J.Mechanisms of human bladder tumor invasion: role of protease cathepsin B . J. Urol. , 144 , 798 – 804 ( 1990. ). [DOI] [PubMed] [Google Scholar]
  • 18. ) Redwood , S. M. , Lie , B. C.‐S. , Weiss , R. E. , Hodge , D. E. and Droller , M. J.Abrogation of the invasion of human bladder tumor cells by using protease inhibitor(s) . Cancer , 69 , 1212 – 1219 ( 1992. ). [DOI] [PubMed] [Google Scholar]
  • 19. ) Laemmli , U. K.Cleavage of structural proteins during the assembly of the head of bacteriophage T4 . Nature , 227 , 680 – 685 ( 1970. ). [DOI] [PubMed] [Google Scholar]
  • 20. ) Schmitt , M. , Goretzki , L. , Janicke , F. , Calvete , J. , Eulitz , M. , Kobayashi , H. , Chucholowski , N. and Graeff , H.Biological and clinical relevance of the urokinase‐type plasminogen activator (uPA) in breast cancer . Biomed. Biochem. Ada , 50 , 4 – 6 , 731 – 741 ( 1991. ). [PubMed] [Google Scholar]
  • 21. ) Còhen , R. L. , Xi , X‐P. , Crowley , C. W. , Lucas , B. K. , Levinson , A. D. and Scuman , M. A.Effects of urokinase receptor occupancy on plasmin generation and proteolysis of basement membrane by human tumor cells . Blood , 78 , 479 – 487 ( 1991. ). [PubMed] [Google Scholar]
  • 22. ) Ossowski , L.In vivo invasion of modified chorioallantoic membrane by tumor cells: the role of cell surface‐bound urokinase . J. Cell Biol. , 107 , 2437 – 2445 ( 1988. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 23. ) Ossowski , L. and Reich , E.Antibodies to plasminogen activator inhibit human tumor metastasis . Cell , 35 , 611 – 619 ( 1983. ). [DOI] [PubMed] [Google Scholar]
  • 24. ) Ossowski , L. and Reich , E.Experimental model for quantitative study of metastasis . Cancer Res. , 40 , 2300 – 2309 ( 1980. ). [PubMed] [Google Scholar]
  • 25. ) Mignatti , P. , Robbins , E. and Rifkin , D. B.Tumor invasion through the human amniotic membrane: requirement for a proteinase cascade . Cell , 47 , 489 – 498 ( 1986. ). [DOI] [PubMed] [Google Scholar]
  • 26. ) Reich , R. , Thompson , E. W. , Iwamoto , Y. , Martin , G. R. , Deason , J. R. , Fuller , G. C. and Miskin , R.Effects of inhibitors of plasminogen activator, serine proteases, and collagenase IV on the invasion of basement membranes by metastatic cells . Cancer Res. , 48 , 3307 – 3312 ( 1988. ). [PubMed] [Google Scholar]
  • 27. ) Hearing , V. J. , Law , L. W. , Corti , A. , Appella , E. and Blasi , F.Modulation of metastatic potential by cell surface urokinase of murine melanoma cells . Cancer Res. , 48 , 1270 – 1278 ( 1988. ). [PubMed] [Google Scholar]
  • 28. ) Cajot , J. F. , Schleuning , W. D. , Medcalf , R. L. , Bamat , J. , Testuz , J. , Liebermann , L. and Sordat , B.Mouse L cells expressing human prourokinase‐type plasminogen activator: effects on extracellular matrix degradation and invasion . J. Cell Biol. , 109 , 915 – 925 ( 1989. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 29. ) Schlechte , W. , Murano , G. and Boyd , D.Examination of the role of the urokinase receptor in human colon cancer mediated laminin degradation . Cancer Res. , 49 , 6064 – 6069 ( 1989. ). [PubMed] [Google Scholar]
  • 30. ) Liotta , L. A. , Tryggvason , K. , Garbisa , S. , Hart , I. , Foltz , C. M. and Shafie , S.Metastatic potential correlates with enzymatic degradation of basement membrane collagen . Nature , 284 , 67 – 68 ( 1980. ). [DOI] [PubMed] [Google Scholar]
  • 31. ) Nakajima , M. , Welch , D. R. , Belloni , P. N. and Nicolson , G. L.Degradation of basement membrane type IV collagen and lung subendothelial matrix by rat mammary adenocarcinoma cell clones of differing metastatic potentials . Cancer Res. , 47 , 4846 – 4876 ( 1987. ). [PubMed] [Google Scholar]
  • 32. ) Sugimura , M. , Kobayashi , H. , Kanayama , N. and Terao , T.Clinical significance of urokinase‐type plasminogen activator (uPA) in invasive cervical cancer of the uterus . Gynecol. Oncol. , 46 , 330 – 336 ( 1992. ). [DOI] [PubMed] [Google Scholar]
  • 33. ) Grondahl‐Hansen , J. , Ralfkiaer , E. , Kirkeby , L. T. , Kristensen , P. , Lund , L. R. and Dano , K.Localization of urokinase‐type plasminogen activator in stromal cells in adenocarcinomas of the colon in humans . Am. J. Pathol. , 138 , 111 – 117 ( 1991. ). [PMC free article] [PubMed] [Google Scholar]
  • 34. ) Pyke , C. , Kristensen , P. , Ralfkiaer , E. , Grondahl‐Hansen , J. , Eriksen , J. , Blasi , F. and Dano , 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. , 138 , 1059 – 1067 ( 1991. ). [PMC free article] [PubMed] [Google Scholar]
  • 35. ) Schmitt , M. , Janicke , F. and Graeff , H.Tumor‐associated fibrinolysis: the prognostic relevance of plasminogen activators uPA and tPA in human breast cancer . Blood Coagulation Fibrinolysis , 1 , 695 – 702 ( 1990. ). [PubMed] [Google Scholar]
  • 36. ) Jänicke , P. , Schmitt , M. , Ulm , K. , Gossner , W. and Graeff , H.Urokinase‐type plasminogen activator antigen and early relapse in breast cancer . Lancet , ii , 1049 ( 1989. ). [DOI] [PubMed] [Google Scholar]
  • 37. ) Liotta , L. A. and Stetler‐Stevenson , W. G.Tumor invasion and metastasis: an imbalance of positive and negative regulation . Cancer Res. ( suppl. ), 51 , 5054s – 5059s ( 1991. ). [PubMed] [Google Scholar]
  • 38. ) Albini , A. , Melchiori , A. , Santi , L. , Liotta , L. A. , Brown , P. D. and Stetler‐Stevenson , W. G.Tumor cell invasion inhibited by TIMP‐2 . J. Natl. Cancer Inst. , 83 , 775 – 779 ( 1991. ). [DOI] [PubMed] [Google Scholar]
  • 39. ) Stephens , R. W. , Pollanen , J. , Tapiovaara , H. , Leung , K.‐C. , Sim , P.‐S. , Salonen , E.‐M. , Ronne , E. , Behrendt , N. , Dano , K. and Vaheri , A.Activation of pro‐urokinase and plasminogen on human sarcoma cells: a proteolytic system with surface‐bound reactions . J. Cell. Biol. , 108 , 1987 – 1995 ( 1989. ). [DOI] [PMC free article] [PubMed] [Google Scholar]

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