Skip to main content
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1987 Dec 1;105(6):2535–2541. doi: 10.1083/jcb.105.6.2535

Urokinase-type plasminogen activator is induced in migrating capillary endothelial cells

PMCID: PMC2114714  PMID: 3121633

Abstract

Cellular migration is an essential component of invasive biological processes, many of which have been correlated with an increase in plasminogen activator production. Endothelial cell migration occurs in vivo during repair of vascular lesions and angiogenesis, and can be induced in vitro by wounding a confluent monolayer of cells. By combining the wounded monolayer model with a substrate overlay technique, we show that cells migrating from the edges of an experimental wound display an increase in urokinase-type plasminogen activator (uPA) activity, and that this activity reverts to background levels upon cessation of movement, when the wound has closed. Our results demonstrate a direct temporal relationship between endothelial cell migration and uPA activity, and suggest that induction of uPA activity is a component of the migratory process.

Full Text

The Full Text of this article is available as a PDF (3.2 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aggeler J., Kapp L. N., Tseng S. C., Werb Z. Regulation of protein secretion in Chinese hamster ovary cells by cell cycle position and cell density. Plasminogen activator, procollagen fibronectin. Exp Cell Res. 1982 Jun;139(2):275–283. doi: 10.1016/0014-4827(82)90252-x. [DOI] [PubMed] [Google Scholar]
  2. Ausprunk D. H., Folkman J. Migration and proliferation of endothelial cells in preformed and newly formed blood vessels during tumor angiogenesis. Microvasc Res. 1977 Jul;14(1):53–65. doi: 10.1016/0026-2862(77)90141-8. [DOI] [PubMed] [Google Scholar]
  3. Bissell M. J., Hall H. G., Parry G. How does the extracellular matrix direct gene expression? J Theor Biol. 1982 Nov 7;99(1):31–68. doi: 10.1016/0022-5193(82)90388-5. [DOI] [PubMed] [Google Scholar]
  4. Bürk R. R. A factor from a transformed cell line that affects cell migration. Proc Natl Acad Sci U S A. 1973 Feb;70(2):369–372. doi: 10.1073/pnas.70.2.369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Danø K., Andreasen P. A., Grøndahl-Hansen J., Kristensen P., Nielsen L. S., Skriver L. Plasminogen activators, tissue degradation, and cancer. Adv Cancer Res. 1985;44:139–266. doi: 10.1016/s0065-230x(08)60028-7. [DOI] [PubMed] [Google Scholar]
  6. Deutsch D. G., Mertz E. T. Plasminogen: purification from human plasma by affinity chromatography. Science. 1970 Dec 4;170(3962):1095–1096. doi: 10.1126/science.170.3962.1095. [DOI] [PubMed] [Google Scholar]
  7. Fishman J. A., Ryan G. B., Karnovsky M. J. Endothelial regeneration in the rat carotid artery and the significance of endothelial denudation in the pathogenesis of myointimal thickening. Lab Invest. 1975 Mar;32(3):339–351. [PubMed] [Google Scholar]
  8. Folkman J. Tumor angiogenesis. Adv Cancer Res. 1985;43:175–203. doi: 10.1016/s0065-230x(08)60946-x. [DOI] [PubMed] [Google Scholar]
  9. Furie M. B., Cramer E. B., Naprstek B. L., Silverstein S. C. Cultured endothelial cell monolayers that restrict the transendothelial passage of macromolecules and electrical current. J Cell Biol. 1984 Mar;98(3):1033–1041. doi: 10.1083/jcb.98.3.1033. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gabbiani G., Gabbiani F., Heimark R. L., Schwartz S. M. Organization of actin cytoskeleton during early endothelial regeneration in vitro. J Cell Sci. 1984 Mar;66:39–50. doi: 10.1242/jcs.66.1.39. [DOI] [PubMed] [Google Scholar]
  11. Gotlieb A. I., May L. M., Subrahmanyan L., Kalnins V. I. Distribution of microtubule organizing centers in migrating sheets of endothelial cells. J Cell Biol. 1981 Nov;91(2 Pt 1):589–594. doi: 10.1083/jcb.91.2.589. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Gotlieb A. I., Spector W. Migration into an in vitro experimental wound: a comparison of porcine aortic endothelial and smooth muscle cells and the effect of culture irradiation. Am J Pathol. 1981 May;103(2):271–282. [PMC free article] [PubMed] [Google Scholar]
  13. Gotlieb A. I., Spector W., Wong M. K., Lacey C. In vitro reendothelialization. Microfilament bundle reorganization in migrating porcine endothelial cells. Arteriosclerosis. 1984 Mar-Apr;4(2):91–96. doi: 10.1161/01.atv.4.2.91. [DOI] [PubMed] [Google Scholar]
  14. Gotlieb A. I., Subrahmanyan L., Kalnins V. I. Microtubule-organizing centers and cell migration: effect of inhibition of migration and microtubule disruption in endothelial cells. J Cell Biol. 1983 May;96(5):1266–1272. doi: 10.1083/jcb.96.5.1266. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Grimaldi G., Di Fiore P., Locatelli E. K., Falco J., Blasi F. Modulation of urokinase plasminogen activator gene expression during the transition from quiescent to proliferative state in normal mouse cells. EMBO J. 1986 May;5(5):855–861. doi: 10.1002/j.1460-2075.1986.tb04295.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Gross J. L., Moscatelli D., Jaffe E. A., Rifkin D. B. Plasminogen activator and collagenase production by cultured capillary endothelial cells. J Cell Biol. 1982 Dec;95(3):974–981. doi: 10.1083/jcb.95.3.974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Gross J. L., Moscatelli D., Rifkin D. B. Increased capillary endothelial cell protease activity in response to angiogenic stimuli in vitro. Proc Natl Acad Sci U S A. 1983 May;80(9):2623–2627. doi: 10.1073/pnas.80.9.2623. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Haudenschild C. C., Schwartz S. M. Endothelial regeneration. II. Restitution of endothelial continuity. Lab Invest. 1979 Nov;41(5):407–418. [PubMed] [Google Scholar]
  19. Hormia M., Badley R. A., Lehto V. P., Virtanen I. Actomyosin organization in stationary and migrating sheets of cultured human endothelial cells. Exp Cell Res. 1985 Mar;157(1):116–126. doi: 10.1016/0014-4827(85)90156-9. [DOI] [PubMed] [Google Scholar]
  20. IYER V. N., SZYBALSKI W. A MOLECULAR MECHANISM OF MITOMYCIN ACTION: LINKING OF COMPLEMENTARY DNA STRANDS. Proc Natl Acad Sci U S A. 1963 Aug;50:355–362. doi: 10.1073/pnas.50.2.355. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Levin E. G., Loskutoff D. J. Comparative studies of the fibrinolytic activity of cultured vascular cells. Thromb Res. 1979;15(5-6):869–878. doi: 10.1016/0049-3848(79)90195-6. [DOI] [PubMed] [Google Scholar]
  22. Levin E. G., Loskutoff D. J. Cultured bovine endothelial cells produce both urokinase and tissue-type plasminogen activators. J Cell Biol. 1982 Sep;94(3):631–636. doi: 10.1083/jcb.94.3.631. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Loskutoff D. J. Effects of acidified fetal bovine serum on the fibrinolytic activity and growth of cells in culture. J Cell Physiol. 1978 Sep;96(3):361–369. doi: 10.1002/jcp.1040960312. [DOI] [PubMed] [Google Scholar]
  24. Loskutoff D. J., Ny T., Sawdey M., Lawrence D. Fibrinolytic system of cultured endothelial cells: regulation by plasminogen activator inhibitor. J Cell Biochem. 1986;32(4):273–280. doi: 10.1002/jcb.240320404. [DOI] [PubMed] [Google Scholar]
  25. Loskutoff D. J., Paul D. Intracellular plasminogen activator activity in growing and quiescent cells. J Cell Physiol. 1978 Oct;97(1):9–16. doi: 10.1002/jcp.1040970103. [DOI] [PubMed] [Google Scholar]
  26. Madri J. A., Stenn K. S. Aortic endothelial cell migration. I. Matrix requirements and composition. Am J Pathol. 1982 Feb;106(2):180–186. [PMC free article] [PubMed] [Google Scholar]
  27. Mascardo R. N., Sherline P. Insulin and multiplication-stimulating activity induce a very rapid centrosomal orientation response to wounding in endothelial cell monolayers. Diabetes. 1984 Nov;33(11):1099–1105. doi: 10.2337/diab.33.11.1099. [DOI] [PubMed] [Google Scholar]
  28. Mignatti P., Robbins E., Rifkin D. B. Tumor invasion through the human amniotic membrane: requirement for a proteinase cascade. Cell. 1986 Nov 21;47(4):487–498. doi: 10.1016/0092-8674(86)90613-6. [DOI] [PubMed] [Google Scholar]
  29. Miskin R., Reich E. Plasminogen activator: induction of synthesis by DNA damage. Cell. 1980 Jan;19(1):217–224. doi: 10.1016/0092-8674(80)90403-1. [DOI] [PubMed] [Google Scholar]
  30. Montesano R., Orci L. Tumor-promoting phorbol esters induce angiogenesis in vitro. Cell. 1985 Sep;42(2):469–477. doi: 10.1016/0092-8674(85)90104-7. [DOI] [PubMed] [Google Scholar]
  31. Montesano R., Pepper M. S., Vassalli J. D., Orci L. Phorbol ester induces cultured endothelial cells to invade a fibrin matrix in the presence of fibrinolytic inhibitors. J Cell Physiol. 1987 Sep;132(3):509–516. doi: 10.1002/jcp.1041320313. [DOI] [PubMed] [Google Scholar]
  32. Montesano R., Vassalli J. D., Baird A., Guillemin R., Orci L. Basic fibroblast growth factor induces angiogenesis in vitro. Proc Natl Acad Sci U S A. 1986 Oct;83(19):7297–7301. doi: 10.1073/pnas.83.19.7297. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Morioka S., Lazarus G. S., Baird J. L., Jensen P. J. Migrating keratinocytes express urokinase-type plasminogen activator. J Invest Dermatol. 1987 Apr;88(4):418–423. doi: 10.1111/1523-1747.ep12469754. [DOI] [PubMed] [Google Scholar]
  34. Moscatelli D., Jaffe E., Rifkin D. B. Tetradecanoyl phorbol acetate stimulates latent collagenase production by cultured human endothelial cells. Cell. 1980 Jun;20(2):343–351. doi: 10.1016/0092-8674(80)90620-0. [DOI] [PubMed] [Google Scholar]
  35. Moscatelli D., Presta M., Joseph-Silverstein J., Rifkin D. B. Both normal and tumor cells produce basic fibroblast growth factor. J Cell Physiol. 1986 Nov;129(2):273–276. doi: 10.1002/jcp.1041290220. [DOI] [PubMed] [Google Scholar]
  36. Moscatelli D., Presta M., Rifkin D. B. Purification of a factor from human placenta that stimulates capillary endothelial cell protease production, DNA synthesis, and migration. Proc Natl Acad Sci U S A. 1986 Apr;83(7):2091–2095. doi: 10.1073/pnas.83.7.2091. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Moscatelli D. Urokinase-type and tissue-type plasminogen activators have different distributions in cultured bovine capillary endothelial cells. J Cell Biochem. 1986;30(1):19–29. doi: 10.1002/jcb.240300104. [DOI] [PubMed] [Google Scholar]
  38. Mullins D. E., Rohrlich S. T. The role of proteinases in cellular invasiveness. Biochim Biophys Acta. 1983 Dec 29;695(3-4):177–214. doi: 10.1016/0304-419x(83)90011-2. [DOI] [PubMed] [Google Scholar]
  39. Ossowski L., Quigley J. P., Kellerman G. M., Reich E. Fibrinolysis associated with oncogenic transformation. Requirement of plasminogen for correlated changes in cellular morphology, colony formation in agar, and cell migration. J Exp Med. 1973 Nov 1;138(5):1056–1064. doi: 10.1084/jem.138.5.1056. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Ossowski L., Reich E. Antibodies to plasminogen activator inhibit human tumor metastasis. Cell. 1983 Dec;35(3 Pt 2):611–619. doi: 10.1016/0092-8674(83)90093-4. [DOI] [PubMed] [Google Scholar]
  41. Pratt B. M., Harris A. S., Morrow J. S., Madri J. A. Mechanisms of cytoskeletal regulation. Modulation of aortic endothelial cell spectrin by the extracellular matrix. Am J Pathol. 1984 Dec;117(3):349–354. [PMC free article] [PubMed] [Google Scholar]
  42. Rohrlich S. T., Rifkin D. B. Patterns of plasminogen activator production in cultured normal embryonic cells. J Cell Biol. 1977 Oct;75(1):31–42. doi: 10.1083/jcb.75.1.31. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Ryan U. S., Absher M., Olazabal B. M., Brown L. M., Ryan J. W. Proliferation of pulmonary endothelial cells: time-lapse cinematography of growth to confluence and restitution of monolayer after wounding. Tissue Cell. 1982;14(4):637–649. doi: 10.1016/0040-8166(82)90054-4. [DOI] [PubMed] [Google Scholar]
  44. Saksela O. Plasminogen activation and regulation of pericellular proteolysis. Biochim Biophys Acta. 1985 Nov 12;823(1):35–65. doi: 10.1016/0304-419x(85)90014-9. [DOI] [PubMed] [Google Scholar]
  45. Schleef R. R., Birdwell C. R. The effect of proteases on endothelial cell migration in vitro. Exp Cell Res. 1982 Oct;141(2):503–508. doi: 10.1016/0014-4827(82)90244-0. [DOI] [PubMed] [Google Scholar]
  46. Schwartz S. M., Haudenschild C. C., Eddy E. M. Endothelial regneration. I. Quantitative analysis of initial stages of endothelial regeneration in rat aortic intima. Lab Invest. 1978 May;38(5):568–580. [PubMed] [Google Scholar]
  47. Scott F. M., Sator de Serrano V., Castellino F. J. Appearance of plasminogen activator activity during a synchronous cycle of a rat adenocarcinoma cell line, PA-III. Exp Cell Res. 1987 Mar;169(1):39–46. doi: 10.1016/0014-4827(87)90222-9. [DOI] [PubMed] [Google Scholar]
  48. Seifter J. L., Aronson P. S. Properties and physiologic roles of the plasma membrane sodium-hydrogen exchanger. J Clin Invest. 1986 Oct;78(4):859–864. doi: 10.1172/JCI112671. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Selden S. C., 3rd, Rabinovitch P. S., Schwartz S. M. Effects of cytoskeletal disrupting agents on replication of bovine endothelium. J Cell Physiol. 1981 Aug;108(2):195–211. doi: 10.1002/jcp.1041080210. [DOI] [PubMed] [Google Scholar]
  50. Selden S. C., 3rd, Schwartz S. M. Cytochalasin B inhibition of endothelial proliferation at wound edges in vitro. J Cell Biol. 1979 May;81(2):348–354. doi: 10.1083/jcb.81.2.348. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Sholley M. M., Gimbrone M. A., Jr, Cotran R. S. Cellular migration and replication in endothelial regeneration: a study using irradiated endothelial cultures. Lab Invest. 1977 Jan;36(1):18–25. [PubMed] [Google Scholar]
  52. Tomasz M., Lipman R., Chowdary D., Pawlak J., Verdine G. L., Nakanishi K. Isolation and structure of a covalent cross-link adduct between mitomycin C and DNA. Science. 1987 Mar 6;235(4793):1204–1208. doi: 10.1126/science.3103215. [DOI] [PubMed] [Google Scholar]
  53. Vassalli J. D., Belin D. Amiloride selectively inhibits the urokinase-type plasminogen activator. FEBS Lett. 1987 Apr 6;214(1):187–191. doi: 10.1016/0014-5793(87)80039-x. [DOI] [PubMed] [Google Scholar]
  54. Vassalli J. D., Dayer J. M., Wohlwend A., Belin D. Concomitant secretion of prourokinase and of a plasminogen activator-specific inhibitor by cultured human monocytes-macrophages. J Exp Med. 1984 Jun 1;159(6):1653–1668. doi: 10.1084/jem.159.6.1653. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Vassalli J. D., Hamilton J., Reich E. Macrophage plasminogen activator: induction by concanavalin A and phorbol myristate acetate. Cell. 1977 Jul;11(3):695–705. doi: 10.1016/0092-8674(77)90086-1. [DOI] [PubMed] [Google Scholar]
  56. Young W. C., Herman I. M. Extracellular matrix modulation of endothelial cell shape and motility following injury in vitro. J Cell Sci. 1985 Feb;73:19–32. doi: 10.1242/jcs.73.1.19. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES