Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1991 May 2;113(4):857–865. doi: 10.1083/jcb.113.4.857

The inhibition of EGF-dependent proliferation of keratinocytes by tyrphostin tyrosine kinase blockers

PMCID: PMC2288989  PMID: 1709168

Abstract

Protein tyrosine kinase blockers of the tyrphostin family inhibited the EGF-dependent proliferation of human and guinea pig keratinocytes grown in culture and induced their growth arrest. These blockers also significantly inhibited the growth of epidermal keratinocytes, but not of dermal cells, in whole skin organ culture from both guinea pig and human origin. The antiproliferative activity of these tyrphostins correlated quantitatively with their potency as inhibitors of EGF receptor autophosphorylation and the EGF-dependent protein phosphorylation of intracellular target proteins in the keratinocyte. Furthermore, no significant cell cytotoxicity or reduction in serine and threonine phosphorylation of many intracellular polypeptides were observed upon incubation of the cells with tyrphostins like AG213. The complete growth arrest induced by the tyrphostins is fully reversible and upon their removal the keratinocytes resumed their growth with the original growth rate. Because of the nontoxic nature of these compounds and their growth-arresting properties, we suggest their use as agents to treat hyperproliferative conditions of human skin.

Full Text

The Full Text of this article is available as a PDF (1.9 MB).

Selected References

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

  1. Barrandon Y., Green H. Cell migration is essential for sustained growth of keratinocyte colonies: the roles of transforming growth factor-alpha and epidermal growth factor. Cell. 1987 Sep 25;50(7):1131–1137. doi: 10.1016/0092-8674(87)90179-6. [DOI] [PubMed] [Google Scholar]
  2. Basler K., Hafen E. Control of photoreceptor cell fate by the sevenless protein requires a functional tyrosine kinase domain. Cell. 1988 Jul 29;54(3):299–311. doi: 10.1016/0092-8674(88)90193-6. [DOI] [PubMed] [Google Scholar]
  3. Bishop J. M. The molecular genetics of cancer. Science. 1987 Jan 16;235(4786):305–311. doi: 10.1126/science.3541204. [DOI] [PubMed] [Google Scholar]
  4. Coffey R. J., Jr, Derynck R., Wilcox J. N., Bringman T. S., Goustin A. S., Moses H. L., Pittelkow M. R. Production and auto-induction of transforming growth factor-alpha in human keratinocytes. 1987 Aug 27-Sep 2Nature. 328(6133):817–820. doi: 10.1038/328817a0. [DOI] [PubMed] [Google Scholar]
  5. Coffey R. J., Jr, Sipes N. J., Bascom C. C., Graves-Deal R., Pennington C. Y., Weissman B. E., Moses H. L. Growth modulation of mouse keratinocytes by transforming growth factors. Cancer Res. 1988 Mar 15;48(6):1596–1602. [PubMed] [Google Scholar]
  6. Derynck R., Roberts A. B., Winkler M. E., Chen E. Y., Goeddel D. V. Human transforming growth factor-alpha: precursor structure and expression in E. coli. Cell. 1984 Aug;38(1):287–297. doi: 10.1016/0092-8674(84)90550-6. [DOI] [PubMed] [Google Scholar]
  7. Elder J. T., Fisher G. J., Lindquist P. B., Bennett G. L., Pittelkow M. R., Coffey R. J., Jr, Ellingsworth L., Derynck R., Voorhees J. J. Overexpression of transforming growth factor alpha in psoriatic epidermis. Science. 1989 Feb 10;243(4892):811–814. doi: 10.1126/science.2916128. [DOI] [PubMed] [Google Scholar]
  8. Filvaroff E., Stern D. F., Dotto G. P. Tyrosine phosphorylation is an early and specific event involved in primary keratinocyte differentiation. Mol Cell Biol. 1990 Mar;10(3):1164–1173. doi: 10.1128/mcb.10.3.1164. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gazit A., Yaish P., Gilon C., Levitzki A. Tyrphostins I: synthesis and biological activity of protein tyrosine kinase inhibitors. J Med Chem. 1989 Oct;32(10):2344–2352. doi: 10.1021/jm00130a020. [DOI] [PubMed] [Google Scholar]
  10. Green H. The keratinocyte as differentiated cell type. Harvey Lect. 1980;74:101–139. [PubMed] [Google Scholar]
  11. Hunter T., Cooper J. A. Protein-tyrosine kinases. Annu Rev Biochem. 1985;54:897–930. doi: 10.1146/annurev.bi.54.070185.004341. [DOI] [PubMed] [Google Scholar]
  12. King L. E., Jr, Gates R. E., Stoscheck C. M., Nanney L. B. The EGF/TGF alpha receptor in skin. J Invest Dermatol. 1990 Jun;94(6 Suppl):164S–170S. doi: 10.1111/1523-1747.ep12876141. [DOI] [PubMed] [Google Scholar]
  13. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  14. Lyall R. M., Zilberstein A., Gazit A., Gilon C., Levitzki A., Schlessinger J. Tyrphostins inhibit epidermal growth factor (EGF)-receptor tyrosine kinase activity in living cells and EGF-stimulated cell proliferation. J Biol Chem. 1989 Aug 25;264(24):14503–14509. [PubMed] [Google Scholar]
  15. Maciag T., Nemore R. E., Weinstein R., Gilchrest B. A. An endocrine approach to the control of epidermal growth: serum-free cultivation of human keratinocytes. Science. 1981 Mar 27;211(4489):1452–1454. doi: 10.1126/science.6970413. [DOI] [PubMed] [Google Scholar]
  16. Margolis B., Rhee S. G., Felder S., Mervic M., Lyall R., Levitzki A., Ullrich A., Zilberstein A., Schlessinger J. EGF induces tyrosine phosphorylation of phospholipase C-II: a potential mechanism for EGF receptor signaling. Cell. 1989 Jun 30;57(7):1101–1107. doi: 10.1016/0092-8674(89)90047-0. [DOI] [PubMed] [Google Scholar]
  17. Posner I., Gazit A., Gilon C., Levitzki A. Tyrphostins inhibit the epidermal growth factor receptor-mediated breakdown of phosphoinositides. FEBS Lett. 1989 Nov 6;257(2):287–291. doi: 10.1016/0014-5793(89)81554-6. [DOI] [PubMed] [Google Scholar]
  18. Rheinwald J. G., Green H. Epidermal growth factor and the multiplication of cultured human epidermal keratinocytes. Nature. 1977 Feb 3;265(5593):421–424. doi: 10.1038/265421a0. [DOI] [PubMed] [Google Scholar]
  19. Rheinwald J. G. Serial cultivation of normal human epidermal keratinocytes. Methods Cell Biol. 1980;21A:229–254. doi: 10.1016/s0091-679x(08)60769-4. [DOI] [PubMed] [Google Scholar]
  20. Ross R. The pathogenesis of atherosclerosis--an update. N Engl J Med. 1986 Feb 20;314(8):488–500. doi: 10.1056/NEJM198602203140806. [DOI] [PubMed] [Google Scholar]
  21. Shechter Y., Yaish P., Chorev M., Gilon C., Braun S., Levitzki A. Inhibition of insulin-dependent lipogenesis and anti-lipolysis by protein tyrosine kinase inhibitors. EMBO J. 1989 Jun;8(6):1671–1676. doi: 10.1002/j.1460-2075.1989.tb03558.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Todaro G. J., Fryling C., De Larco J. E. Transforming growth factors produced by certain human tumor cells: polypeptides that interact with epidermal growth factor receptors. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5258–5262. doi: 10.1073/pnas.77.9.5258. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Turbitt M. L., Akhurst R. J., White S. I., MacKie R. M. Localization of elevated transforming growth factor-alpha in psoriatic epidermis. J Invest Dermatol. 1990 Aug;95(2):229–232. doi: 10.1111/1523-1747.ep12478077. [DOI] [PubMed] [Google Scholar]
  24. Ullrich A., Schlessinger J. Signal transduction by receptors with tyrosine kinase activity. Cell. 1990 Apr 20;61(2):203–212. doi: 10.1016/0092-8674(90)90801-k. [DOI] [PubMed] [Google Scholar]
  25. Weissman B., Aaronson S. A. Members of the src and ras oncogene families supplant the epidermal growth factor requirement of BALB/MK-2 keratinocytes and induce distinct alterations in their terminal differentiation program. Mol Cell Biol. 1985 Dec;5(12):3386–3396. doi: 10.1128/mcb.5.12.3386. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Wille J. J., Jr, Pittelkow M. R., Shipley G. D., Scott R. E. Integrated control of growth and differentiation of normal human prokeratinocytes cultured in serum-free medium: clonal analyses, growth kinetics, and cell cycle studies. J Cell Physiol. 1984 Oct;121(1):31–44. doi: 10.1002/jcp.1041210106. [DOI] [PubMed] [Google Scholar]
  27. Yaish P., Gazit A., Gilon C., Levitzki A. Blocking of EGF-dependent cell proliferation by EGF receptor kinase inhibitors. Science. 1988 Nov 11;242(4880):933–935. doi: 10.1126/science.3263702. [DOI] [PubMed] [Google Scholar]
  28. Yarden Y., Ullrich A. Growth factor receptor tyrosine kinases. Annu Rev Biochem. 1988;57:443–478. doi: 10.1146/annurev.bi.57.070188.002303. [DOI] [PubMed] [Google Scholar]

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

RESOURCES