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. 1984 Jan;4(1):160–165. doi: 10.1128/mcb.4.1.160

Modulation of cell-associated plasminogen activator activity by cocultivation of a stem cell and its tumorigenic descendant.

H Y Liu, P P Yang, D L Toledo, W F Mangel
PMCID: PMC368670  PMID: 6538259

Abstract

The effect of the presence of one cell type on the plasminogen activator activity of another cell type was studied. The cell types, AC and D, were isolated from a rat neuroblastoma (I. Imada and N. Sueoka, Dev. Biol. 66:97-108, 1978). AC cells are stem cells capable of multipotential differentiation in vitro and have little or no cell-associated plasminogen activator activity. D cells are tumorigenic and have high levels of cell-associated plasminogen activator activity. When AC cells were cocultivated with D cells, the plasminogen activator activity of the D cells was dramatically inhibited. The presence of as few as 1,250 AC cells inhibited 70% of the plasminogen activator activity of 20,000 D cells, as determined by a highly quantitative assay. The amount of inhibition by AC cells was proportional to the number of AC cells present. At increasing numbers of AC cells and a constant number of D cells, the Vmax for the activation of plasminogen proportionately decreased and the Km remained constant, implying that AC cells did not alter the structure or concentration of plasminogen. Inhibition was not mediated by a soluble inhibitor secreted by AC cells. Rather, attachment of AC cells adjacent to D cells, i.e., cell-to-cell contact, seemed to be required for inhibition. The substratum-attached material of AC cells, that which remained on the microwell surface after removal of AC cells with EDTA, inhibited D cell plasminogen activator activity. If plasminogen activator activity is involved in metastasis, then regulation of the plasminogen activator activity of one cell type by another cell type may be involved in determining which cells in a tumor can metastasize and where secondary tumors can arise.

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Selected References

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

  1. BACHMANN F., FLETCHER A. P., ALKJAERSIG N., SHERRY S. PARTIAL PURIFICATION AND PROPERTIES OF THE PLASMINOGEN ACTIVATOR FROM PIG HEART. Biochemistry. 1964 Oct;3:1578–1585. doi: 10.1021/bi00898a033. [DOI] [PubMed] [Google Scholar]
  2. Beers W. H., Strickland S. A cell culture assay for follicle-stimulating hormone. J Biol Chem. 1978 Jun 10;253(11):3877–3881. [PubMed] [Google Scholar]
  3. Bernik M. B., Kwaan H. C. Origin of fibrinolytic activity in cultures of the human kidney. J Lab Clin Med. 1967 Oct;70(4):650–661. [PubMed] [Google Scholar]
  4. Bernik M. B., Kwaan H. C. Plasminogen activator activity in cultures from human tissues. An immunological and histochemical study. J Clin Invest. 1969 Sep;48(9):1740–1753. doi: 10.1172/JCI106140. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Castellino F. J., Sodetz J. M. Rabbit plasminogen and plasmin isozymes. Methods Enzymol. 1976;45:273–286. doi: 10.1016/s0076-6879(76)45026-7. [DOI] [PubMed] [Google Scholar]
  6. Christman J. K., Silagi S., Newcomb E. W., Silverstein S. C., Acs G. Correlated suppression by 5-bromodeoxyuridine of tumorigenicity and plasminogen activator in mouse melanoma cells. Proc Natl Acad Sci U S A. 1975 Jan;72(1):47–50. doi: 10.1073/pnas.72.1.47. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cleland W. W. The statistical analysis of enzyme kinetic data. Adv Enzymol Relat Areas Mol Biol. 1967;29:1–32. doi: 10.1002/9780470122747.ch1. [DOI] [PubMed] [Google Scholar]
  8. Gallimore P. H., McDougall J. K., Chen L. B. In vitro traits of adenovirus-transformed cell lines and their relevance to tumorigenicity in nude mice. Cell. 1977 Apr;10(4):669–678. doi: 10.1016/0092-8674(77)90100-3. [DOI] [PubMed] [Google Scholar]
  9. Imada M., Sueoka N. Clonal sublines of rat neurotumor RT4 and cell differentiation. I. Isolation and characterization of cell lines and cell type conversion. Dev Biol. 1978 Sep;66(1):97–108. doi: 10.1016/0012-1606(78)90276-2. [DOI] [PubMed] [Google Scholar]
  10. Imada M., Sueoka N., Rifkin D. B. Clonal sublines of rat neurotumor RT4 and cell differentiation. II. A conversion coupling of tumorigenicity and a glial property. Dev Biol. 1978 Sep;66(1):109–116. doi: 10.1016/0012-1606(78)90277-4. [DOI] [PubMed] [Google Scholar]
  11. Kyner D., Christman J., Acs G., Silagi S., Newcomb E. W., Silverstein S. C. Co-cultivation of tumorigenic mouse melanoma cells with cells of a non-tumorigenic subclone inhibits plasminogen activator expression by the melanoma cells. J Cell Physiol. 1978 May;95(2):159–167. doi: 10.1002/jcp.1040950205. [DOI] [PubMed] [Google Scholar]
  12. Laterra J., Norton E. K., Izzard C. S., Culp L. A. Contact formation by fibroblasts adhering to heparan sulfate-binding substrata (fibronectin or platelet factor 4). Exp Cell Res. 1983 Jun;146(1):15–27. doi: 10.1016/0014-4827(83)90320-8. [DOI] [PubMed] [Google Scholar]
  13. Laterra J., Silbert J. E., Culp L. A. Cell surface heparan sulfate mediates some adhesive responses to glycosaminoglycan-binding matrices, including fibronectin. J Cell Biol. 1983 Jan;96(1):112–123. doi: 10.1083/jcb.96.1.112. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Lawrence T. S., Beers W. H., Gilula N. B. Transmission of hormonal stimulation by cell-to-cell communication. Nature. 1978 Apr 6;272(5653):501–506. doi: 10.1038/272501a0. [DOI] [PubMed] [Google Scholar]
  15. Leytus S. P., Peltz G. A., Liu H. Y., Cannon J. F., Peltz S. W., Livingston D. C., Brocklehurst J. R., Mangel W. F. A quantitative assay for the activation of plasminogen by transformed cells in situ and by urokinase. Biochemistry. 1981 Jul 21;20(15):4307–4314. doi: 10.1021/bi00518a011. [DOI] [PubMed] [Google Scholar]
  16. Liu H. Y., Peltz G. A., Leytus S. P., Livingston C., Brocklehurst J., Mangel W. F. Sensitive assay for plasminogen activator of transformed cells. Proc Natl Acad Sci U S A. 1980 Jul;77(7):3796–3800. doi: 10.1073/pnas.77.7.3796. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Liu H. Y., Peltz S. W., Mangel W. F. Modulation of the plasminogen activator activity of a transformed cell line by cell density. Mol Cell Biol. 1982 Nov;2(11):1410–1416. doi: 10.1128/mcb.2.11.1410. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Livingston D. C., Brocklehurst J. R., Cannon J. F., Leytus S. P., Wehrly J. A., Peltz S. W., Peltz G. A., Mangel W. F. Synthesis and characterization of a new fluorogenic active-site titrant of serine proteases. Biochemistry. 1981 Jul 21;20(15):4298–4306. doi: 10.1021/bi00518a010. [DOI] [PubMed] [Google Scholar]
  19. Marshall C. J., Franks L. M., Carbonell A. W. Markers of neoplastic transformation in epithelial cell lines derived from human carcinomas. J Natl Cancer Inst. 1977 Jun;58(6):1743–1751. doi: 10.1093/jnci/58.6.1743. [DOI] [PubMed] [Google Scholar]
  20. Newcomb E. W., Silverstein S. C., Silagi S. Malignant mouse melanoma cells do not form tumors when mixed with cells of a non-malignant subclone: relationships between plasminogen activator expression by the tumor cells and the host's immune response. J Cell Physiol. 1978 May;95(2):169–177. doi: 10.1002/jcp.1040950206. [DOI] [PubMed] [Google Scholar]
  21. Nicolson G. L. Cancer metastasis. Organ colonization and the cell-surface properties of malignant cells. Biochim Biophys Acta. 1982 Dec 21;695(2):113–176. doi: 10.1016/0304-419x(82)90020-8. [DOI] [PubMed] [Google Scholar]
  22. Ossowski L., Biegel D., Reich E. Mammary plasminogen activator: correlation with involution, hormonal modulation and comparison between normal and neoplastic tissue. Cell. 1979 Apr;16(4):929–940. doi: 10.1016/0092-8674(79)90108-9. [DOI] [PubMed] [Google Scholar]
  23. 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]
  24. Ossowski L., Reich E. Experimental model for quantitative study of metastasis. Cancer Res. 1980 Jul;40(7):2300–2309. [PubMed] [Google Scholar]
  25. Ossowski L., Reich E. Loss of malignancy during serial passage of human carcinoma in culture and discordance between malignancy and transformation parameters. Cancer Res. 1980 Jul;40(7):2310–2315. [PubMed] [Google Scholar]
  26. Peltz S. W., Hardt T. A., Mangel W. F. Positive regulation of activation of plasminogen by urokinase: differences in Km for (glutamic acid)-plasminogen and lysine-plasminogen and effect of certain alpha, omega-amino acids. Biochemistry. 1982 May 25;21(11):2798–2804. doi: 10.1021/bi00540a035. [DOI] [PubMed] [Google Scholar]
  27. Pollack R., Risser R., Conlon S., Rifkin D. Plasminogen activator production accompanies loss of anchorage regulation in transformation of primary rat embryo cells by simian virus 40. Proc Natl Acad Sci U S A. 1974 Dec;71(12):4792–4796. doi: 10.1073/pnas.71.12.4792. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Simpson I., Rose B., Loewenstein W. R. Size limit of molecules permeating the junctional membrane channels. Science. 1977 Jan 21;195(4275):294–296. doi: 10.1126/science.831276. [DOI] [PubMed] [Google Scholar]
  29. Stiles C. D., Desmond W., Chuman L. M., Sato G., Saier M. H., Jr Relationship of cell growth behavior in vitro to tumorigenicity in athymic nude mice. Cancer Res. 1976 Sep;36(9 PT1):3300–3305. [PubMed] [Google Scholar]
  30. Strickland S., Beers W. H. Studies on the role of plasminogen activator in ovulation. In vitro response of granulosa cells to gonadotropins, cyclic nucleotides, and prostaglandins. J Biol Chem. 1976 Sep 25;251(18):5694–5702. [PubMed] [Google Scholar]
  31. Tomozawa Y., Sueoka N. In vitro segregation of different cell lines with neuronal and glial properties from a stem cell line of rat neurotumor RT4. Proc Natl Acad Sci U S A. 1978 Dec;75(12):6305–6309. doi: 10.1073/pnas.75.12.6305. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Unkeless J. C., Gordon S., Reich E. Secretion of plasminogen activator by stimulated macrophages. J Exp Med. 1974 Apr 1;139(4):834–850. doi: 10.1084/jem.139.4.834. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Unkeless J. C., Tobia A., Ossowski L., Quigley J. P., Rifkin D. B., Reich E. An enzymatic function associated with transformation of fibroblasts by oncogenic viruses. I. Chick embryo fibroblast cultures transformed by avian RNA tumor viruses. J Exp Med. 1973 Jan 1;137(1):85–111. doi: 10.1084/jem.137.1.85. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Wiman B., Collen D. Molecular mechanism of physiological fibrinolysis. Nature. 1978 Apr 6;272(5653):549–550. doi: 10.1038/272549a0. [DOI] [PubMed] [Google Scholar]
  35. Wolf B. A., Goldberg A. R. Rous-sarcoma-virus-transformed fibroblasts having low levels of plasminogen activator. Proc Natl Acad Sci U S A. 1976 Oct;73(10):3613–3617. doi: 10.1073/pnas.73.10.3613. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Yang N. S., Park C., Longley C., Furmanski P. Effect of the extracellular matrix on plasminogen activator isozyme activities of human mammary epithelial cells in culture. Mol Cell Biol. 1983 Jun;3(6):982–990. doi: 10.1128/mcb.3.6.982. [DOI] [PMC free article] [PubMed] [Google Scholar]

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