Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1982 Dec 1;95(3):974–981. doi: 10.1083/jcb.95.3.974

Plasminogen activator and collagenase production by cultured capillary endothelial cells

JL Gross, D Moscatelli, EA Jaffe, DB Rifkin
PMCID: PMC2112903  PMID: 6185506

Abstract

Cultured bovine capillary endothelial (BCE) cells produce low levels of collagenolytic activity and significant amounts of the serine protease plasminogen activator (PA). When grown in the presence of nanomolar quantities of the tumor promoter 12-O-tetradecanoyl phorbol-13-acetate (TPA), BCE cells produced 5-15 times more collagenolytic activity and 2-10 times more PA than untreated cells. The enhanced production of these enzymes was dependent on the dose of TPA used, with maximal response at 10(-7) to 10(-8) M. Phorbol didecanoate (PDD), an analog of TPA which is an active tumor promoter, also increased protease production. 4-O-methyl-TPA and 4α-PDD, two analogs of TPA which are inactive as tumor promoters, had no effect on protease production. Increased PA and collagenase activities were detected within 7.5 and 19 h, respectively, after the addition of TPA. The TPA-stimulated BCE cells synthesized a urokinase-type PA and a typical vertebrate collagenase. BCE cells were compared with bovine aortic endothelial (BAE) cells and bovine embryonic skin (BES) fibroblasts with respect to their production of protease in response to TPA. Under normal growth conditions, low levels of collagenolyic activity were detected in the culture fluids from BCE, BAE, and BES cells. BCE cells produced 5-13 times the basal levels of collagenolytic activity in response to TPA, whereas BAE cells and BES fibroblasts showed a minimal response to TPA. Both BCE and BAE cells exhibited relatively high basal levels of PA, the production of which was stimulated approximately threefold by the addition of TPA. The observation that BCE cells and not BAE cells produced high levels of both PA and collagenase activities in response to TPA demonstrates a significant difference between these two types of endothelial cells and suggests that the enhanced detectable activities are a property unique to bovine capillary and microvessel and endothelial cells.

Full Text

The Full Text of this article is available as a PDF (1.3 MB).

Selected References

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

  1. Astedt B., Barlow G., Holmberg L. Time-related release of various molecular forms of urokinase in tissue culture. Thromb Res. 1977 Aug;11(2):149–153. doi: 10.1016/0049-3848(77)90033-0. [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. Binder B. R., Spragg J., Austen K. F. Purification and characterization of human vascular plasminogen activator derived from blood vessel perfusates. J Biol Chem. 1979 Mar 25;254(6):1998–2003. [PubMed] [Google Scholar]
  4. Davison P. M., Karasek M. A. Human dermal microvascular endothelial cells in vitro: effect of cyclic AMP on cellular morphology and proliferation rate. J Cell Physiol. 1981 Feb;106(2):253–258. doi: 10.1002/jcp.1041060211. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. Folkman J., Cotran R. Relation of vascular proliferation to tumor growth. Int Rev Exp Pathol. 1976;16:207–248. [PubMed] [Google Scholar]
  7. Folkman J., Haudenschild C. C., Zetter B. R. Long-term culture of capillary endothelial cells. Proc Natl Acad Sci U S A. 1979 Oct;76(10):5217–5221. doi: 10.1073/pnas.76.10.5217. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Folkman J., Haudenschild C. Angiogenesis in vitro. Nature. 1980 Dec 11;288(5791):551–556. doi: 10.1038/288551a0. [DOI] [PubMed] [Google Scholar]
  9. Glaser B. M., D'Amore P. A., Michels R. G., Patz A., Fenselau A. Demonstration of vasoproliferative activity from mammalian retina. J Cell Biol. 1980 Feb;84(2):298–304. doi: 10.1083/jcb.84.2.298. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  11. Laug W. E., Tokes Z. A., Benedict W. F., Sorgente N. Anchorage independent growth and plasminogen activator production by bovine endothelial cells. J Cell Biol. 1980 Feb;84(2):281–293. doi: 10.1083/jcb.84.2.281. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. 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]
  14. Levin E. G., Loskutoff D. J. Serum-mediated suppression of cell-associated plasminogen activator activity in cultured endothelial cells. Cell. 1980 Dec;22(3):701–707. doi: 10.1016/0092-8674(80)90546-2. [DOI] [PubMed] [Google Scholar]
  15. Liotta L. A., Tryggvason K., Garbisa S., Hart I., Foltz C. M., Shafie S. Metastatic potential correlates with enzymatic degradation of basement membrane collagen. Nature. 1980 Mar 6;284(5751):67–68. doi: 10.1038/284067a0. [DOI] [PubMed] [Google Scholar]
  16. Loskutoff D. J., Edgington T. E. Synthesis of a fibrinolytic activator and inhibitor by endothelial cells. Proc Natl Acad Sci U S A. 1977 Sep;74(9):3903–3907. doi: 10.1073/pnas.74.9.3903. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. 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]
  18. Unkeless J., Dano K., Kellerman G. M., Reich E. Fibrinolysis associated with oncogenic transformation. Partial purification and characterization of the cell factor, a plasminogen activator. J Biol Chem. 1974 Jul 10;249(13):4295–4305. [PubMed] [Google Scholar]
  19. Wigler M., Weinstein I. B. Tumour promotor induces plasminogen activator. Nature. 1976 Jan 22;259(5540):232–233. doi: 10.1038/259232a0. [DOI] [PubMed] [Google Scholar]
  20. Zetter B. R. Migration of capillary endothelial cells is stimulated by tumour-derived factors. Nature. 1980 May 1;285(5759):41–43. doi: 10.1038/285041a0. [DOI] [PubMed] [Google Scholar]

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

RESOURCES