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Protein kinase C inhibitor calphostin C prevents cytokine-induced angiogenesis in the rat

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Abstract

A rat sponge implant model was used to examine the role of protein kinase C (PKC) in angiogenesis, Neovascular response was determined by measurements of relative sponge blood flow by a133Xe clearance technique and confirmed histologically. Morphometric analysis was used to quantitate the amount of fibrovascular growth in the sponges. Daily doses of recombinant human basic fibroblast growth factor (bFGF, 100 ng), tumor necrosis factor-alpha (TNF-α, 50 ng), or interleukin1-alpha (IL-1α, 50 ng) caused neovascular responses that were blocked by daily coadministration of the selective PKC inhibitor, calphostin C (4μg). To confirm that calphostin C was able to inhibit PKC in vivo, its effect on the angiogenic response elicited by the PKC activator, phorbol 12-myristate 13-acetate (PMA, 30μg) was examined. The blood flow and morphometric data clearly showed that the intense neovascularization induced by PMA was totally suppressed by coadministration of calphostin C (4μg). Thus, these results suggest that cytokine-induced angiogenesis may be mediated in part through the activation of PKC and that selective inhibition of this enzyme could have therapeutic benefit in angiogenic diseases.

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References

  1. Folkman, J., andM. Klagsbrun. 1987. Angiogenic factors.Science 235:442–447.

    PubMed  Google Scholar 

  2. Folkman, J., andY. Shing. 1992. Angiogenesis.J. Biol. Chem. 267:10931–10934.

    PubMed  Google Scholar 

  3. Fan, T.-P. D., andS. Brem. 1992. Angiosuppression.In The Search for New Anti-Cancer Drugs. M. J. Waring and B. A. J. Ponder editors. Cancer Biology Series, Vol. 3, Kluwer Academic Publishers, Lancaster, U.K. 185–229.

    Google Scholar 

  4. Fan, T.-P. D. 1994. Angiosuppressive therapy for cancer.Trends Pharmacol. Sci. 15:33–36.

    PubMed  Google Scholar 

  5. Montesano, R., andL. Orci. 1985. Tumor-promoting phorbol esters induce angiogenesis in vitro.Cell 42:469–477.

    PubMed  Google Scholar 

  6. Doctrow, S. R., andJ. Folkman. 1987. Protein kinase C activators suppress stimulation of capillary endothelial cell growth by angiogenic endothelial mitogens.J. Cell Biol. 104:679–687.

    PubMed  Google Scholar 

  7. Morris, P. B., T. Hida, P. J. Blackshear, G. K. Klintworth, andJ. L. Swain. 1988. Tumor-promoting phorbol esters induce angiogenesis in vivo.Am. J. Physiol. 254:C318-C322.

    PubMed  Google Scholar 

  8. Presta, M., J. A. M. Maier, andG. Ragnotti. 1989. The mitogenic signaling pathway but not the plasminogen activator-inducing pathway of basic fibroblast growth factor is mediated through protein kinase C in fetal bovine aortic endothelial cells.J. Cell Biol. 109:1877–1884.

    PubMed  Google Scholar 

  9. Guesdon, F., N. Freshney, R. J. Waller, L. Rawlinson, andJ. Saklatvala. 1993. Interleukin 1 and tumour necrosis factor stimulate two novel protein kinases that phosphorylate the heat shock protein hsp27 andβ-casein.J. Biol. Chem. 268:4236–4243.

    PubMed  Google Scholar 

  10. Saklatvala, J., L. M. Rawlinson, C. J. Marshall, andM. Kracht. 1993. Interleukin-1 and tumour necrosis factor activate the mitogen activated protein (MAP) kinase kinase in cultured cells.FEBS Lett. 334:189–192.

    PubMed  Google Scholar 

  11. Hu, D. E., Y. Hori, M. Presta, G. A. Gresham, andT.-P. D. Fan. 1994. Inhibition of angiogenesis by IL-1 receptor antagonist and selected cytokine antibodies.Inflammation 18(1):45–58.

    PubMed  Google Scholar 

  12. Hu, D. E., andT.-P. D. Fan. 1993. [Leu8]des-Arg9-bradykinin inhibits the synergistic interaction between bradykinin and interleukin-1 in angiogenesis.Br. J. Pharmacol. 109:14–17.

    PubMed  Google Scholar 

  13. Fan, T.-P. D., D. E. Hu, S. Guard, G. A. Gresham, andK. J. Watling. 1993. Stimulation of angiogenesis by substance P and interleukin-1 in the rat and its inhibition by NK1 or interleukin-1 receptor antagonists.Br. J. Pharmacol. 110:43–49.

    PubMed  Google Scholar 

  14. Kobayashi, E., H. Nakano, M. Morimoto, andT. Tamaoki. 1989. Calphostin C (UCN-1028C), a novel microbial compound, is a highly potent and specific inhibitor of protein kinase C.Biochem. Biophys. Res. Commun. 159:548–553.

    PubMed  Google Scholar 

  15. Andrade, S. P., T.-P. D. Fan, andG. P. Lewis. 1987. Quantitative in-vivo studies on angiogenesis in a rat sponge model.Br. J. Exp. Patho. 68:755–766.

    Google Scholar 

  16. Mayhew, T. M., andA. K. Sharma. 1984. Sampling schemes for estimating nerve fibre size. II. Methods for nerve trunks of mixed fascicularity.J. Anat. 139:45–58.

    PubMed  Google Scholar 

  17. Nishizuka, Y. 1984. The role of protein kinase C in cell surface signal transduction and tumour promotion.Nature 308:693–698.

    PubMed  Google Scholar 

  18. Daviet, I., J. M. Herbert, andJ. P. Maffrand. 1990. Involvement of protein kinase C in the mitogenic and chemotaxis effects of basic fibroblast growth factor on bovine cerebral cortex capillary endothelial cells.FEBS Lett. 259:315–317.

    PubMed  Google Scholar 

  19. Dinarello, C. A. 1991. Interleukin 1 and interleukin 1 antagonism.Blood 77:1627–1652.

    PubMed  Google Scholar 

  20. Hu, D. E., Y. Hori, andT.-P. D. Fan. 1993. Interleukin-8 stimulates angiogenesis in rats.Inflammation 17:135–143.

    PubMed  Google Scholar 

  21. Leibovich, S. J., P. J. Polverini, H. M. Shepard, D. M. Wiseman, V. Shively, andN. Niseir. 1987. Macrophage-induced angiogenesis is mediated by tumour necrosis factor-alpha.Nature 329:630–632.

    PubMed  Google Scholar 

  22. Fajardo, L. F., H. H. Kwan, J. Kowalski, S. D. Prionas, andA. C. Allison. 1992. Dual role of tumor necrosis factor-α in angiogenesis.Am. J. Pathol. 140:539–544.

    PubMed  Google Scholar 

  23. Guy, G. R., S. P. Chua, N. S. Wono, S. B. Ng, andY. H. Tan. 1991. Interleukin 1 and tumour necrosis factor activate common multiple protein kinases in human fibroblasts.J. Biol. Chem. 266:14343–14352.

    PubMed  Google Scholar 

  24. Bird, T. A., andJ. Saklatvala. 1990. Interleukin-1 signal transduction. Increased GTP binding and hydrolysis in membranes of a murine thymoma line (EL4).J. Biol. Chem. 265:235–240.

    PubMed  Google Scholar 

  25. Muegge, K., andS. K. Durum. 1990. Cytokines and transcription factors.Cytokine 2:1–8.

    PubMed  Google Scholar 

  26. Pepper, M. S., D. Belin, R. Montesano, L. Orci, andJ. D. Vassalli. 1990. Transforming growth factor-beta 1 modulates basic fibroblast growth factor-induced proteolytic and angiogenic properties of endothelial cells in vitro.J. Cell Biol. 111:743–755.

    PubMed  Google Scholar 

  27. Niedbala, M. J., andM. S. Picarella. 1993. Role of protein kinase C in tumour necrosis factor induction of endothelial cell urokinase type plasminogen activator.Blood 81:2608–2617.

    PubMed  Google Scholar 

  28. Mischak, H., J. H. Pierce, J. Goodnight, M. Kazanietz, P. M. Blumberg, andJ. F. Mushinski. 1993. Phorbol ester-induced myeloid differentiation is mediated by protein kinaseC-α andδ and not by protein kinase C-βII,-ge, -ζ, and-η.J. Biol. Chem. 268:20110–20115.

    PubMed  Google Scholar 

  29. Wilkinson, S. E., andT. J. Hallam. 1994. Protein kinase C: Is its pivotal role in cellular activation over-stated?Trends Pharmacol. Sci. 15:53–57.

    PubMed  Google Scholar 

  30. Hu, D. E., and T.-P. D.Fan. 1995. Suppression of VEGF-induced angiogenesis by the protein tyrosine kinase inhibitor lavendustin A.Br. J. Pharmacol. (in press).

  31. Fotsis, T., M. Pepper, H. Adlercreutz, G. Fleischmann, T. Hase, R. Montesano, andL. Schweigerer. 1993. Genistein, a dietary-derived inhibitor of in vitro angiogenesis.Proc. Natl. Acad. Sci. U.S.A. 90:2690–2694.

    PubMed  Google Scholar 

  32. Oikawa, T., K. Hirotani, M. Shimamura, H. Ashino-Fuse, andT. Iwaguchi. 1989. Powerful antiangiogenic activity of herbimycin A (named angiostatic antibiotic).J. Antibiot. 42:1202–1204.

    PubMed  Google Scholar 

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  1. Department of Pharmacology, University of Cambridge, Tennis Court Road, CB2 1QJ, Cambridge, UK

    D. E. Hu & T. -P. D. Fan

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Hu, D.E., Fan, T.P.D. Protein kinase C inhibitor calphostin C prevents cytokine-induced angiogenesis in the rat. Inflammation 19, 39–54 (1995). https://doi.org/10.1007/BF01534379

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