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. 1985 Dec;82(23):8053–8056. doi: 10.1073/pnas.82.23.8053

Activation of Na+/H+ exchange in cultured fibroblasts: synergism and antagonism between phorbol ester, Ca2+ ionophore, and growth factors.

L M Vicentini, M L Villereal
PMCID: PMC391440  PMID: 2999790

Abstract

The effects of phorbol 12-myristate 13-acetate (PMA), a potent activator of protein kinase C, on Na+ influx were investigated in cultured human foreskin fibroblasts (HSWP cells). We report here that in serum-deprived HSWP cells the addition of PMA alone has no significant effect on Na+ influx. However, the addition of PMA to cells whose Na+/H+ exchanger is partially activated with a submaximal dose of the Ca2+ ionophore A23187 leads to a larger stimulation than seen with A23187 alone. These data suggest that although stimulation of protein kinase C is not a sufficient signal to activate the Na+/H+ exchanger in HSWP cells or in another human foreskin line (Jackson fibroblasts) studied, there are some cooperative effects of protein kinase C activation with a rise in Ca2+ to stimulate Na+/H+ exchange. In addition, we found that PMA actually inhibits the mitogen-induced stimulation of Na+ influx in HSWP and Jackson fibroblasts. This observation strengthens the argument that in these cells activation of protein kinase C is not sufficient to activate Na+/H+ exchange and suggests that there is a negative feedback control via protein kinase C that inhibits some signal that is necessary for activating Na+/H+ exchange. However, in contrast to observations in HSWP cells, we were able to activate the Na+/H+ exchanger in mouse 3T3 and human WI-38 cells with PMA alone, suggesting that there is some diversity in the mechanism for activation of Na+/H+ exchange in different types of fibroblasts.

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Selected References

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

  1. Davis B. A., Schwartz A., Samaha F. J., Kranias E. G. Regulation of cardiac sarcoplasmic reticulum calcium transport by calcium-calmodulin-dependent phosphorylation. J Biol Chem. 1983 Nov 25;258(22):13587–13591. [PubMed] [Google Scholar]
  2. Dicker P., Rozengurt E. Phorbol ester stimulation of Na influx and Na-K pump activity in Swiss 3T3 cells. Biochem Biophys Res Commun. 1981 May 15;100(1):433–441. doi: 10.1016/s0006-291x(81)80115-5. [DOI] [PubMed] [Google Scholar]
  3. Fain J. N., Li S. Y., Litosch I., Wallace M. Synergistic activation of rat hepatocyte glycogen phosphorylase by A23187 and phorbol ester. Biochem Biophys Res Commun. 1984 Feb 29;119(1):88–94. doi: 10.1016/0006-291x(84)91622-x. [DOI] [PubMed] [Google Scholar]
  4. Friedman B., Frackelton A. R., Jr, Ross A. H., Connors J. M., Fujiki H., Sugimura T., Rosner M. R. Tumor promoters block tyrosine-specific phosphorylation of the epidermal growth factor receptor. Proc Natl Acad Sci U S A. 1984 May;81(10):3034–3038. doi: 10.1073/pnas.81.10.3034. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Hesketh T. R., Moore J. P., Morris J. D., Taylor M. V., Rogers J., Smith G. A., Metcalfe J. C. A common sequence of calcium and pH signals in the mitogenic stimulation of eukaryotic cells. Nature. 1985 Feb 7;313(6002):481–484. doi: 10.1038/313481a0. [DOI] [PubMed] [Google Scholar]
  6. Mix L. L., Dinerstein R. J., Villereal M. L. Mitogens and melittin stimulate an increase in intracellular free calcium concentration in human fibroblasts. Biochem Biophys Res Commun. 1984 Feb 29;119(1):69–75. doi: 10.1016/0006-291x(84)91619-x. [DOI] [PubMed] [Google Scholar]
  7. Moolenaar W. H., Tertoolen L. G., de Laat S. W. Phorbol ester and diacylglycerol mimic growth factors in raising cytoplasmic pH. Nature. 1984 Nov 22;312(5992):371–374. doi: 10.1038/312371a0. [DOI] [PubMed] [Google Scholar]
  8. Moolenaar W. H., Tsien R. Y., van der Saag P. T., de Laat S. W. Na+/H+ exchange and cytoplasmic pH in the action of growth factors in human fibroblasts. Nature. 1983 Aug 18;304(5927):645–648. doi: 10.1038/304645a0. [DOI] [PubMed] [Google Scholar]
  9. Muldoon L. L., Dinerstein R. J., Villereal M. L. Intracellular pH in human fibroblasts: effect of mitogens, A23187, and phospholipase activation. Am J Physiol. 1985 Jul;249(1 Pt 1):C140–C148. doi: 10.1152/ajpcell.1985.249.1.C140. [DOI] [PubMed] [Google Scholar]
  10. Naka M., Nishikawa M., Adelstein R. S., Hidaka H. Phorbol ester-induced activation of human platelets is associated with protein kinase C phosphorylation of myosin light chains. Nature. 1983 Dec 1;306(5942):490–492. doi: 10.1038/306490a0. [DOI] [PubMed] [Google Scholar]
  11. Nishizuka Y. The role of protein kinase C in cell surface signal transduction and tumour promotion. Nature. 1984 Apr 19;308(5961):693–698. doi: 10.1038/308693a0. [DOI] [PubMed] [Google Scholar]
  12. Owen N. E., Villereal M. L. Effect of the intracellular Ca+2 antagonist TMB-8 on serum-stimulated Na+ influx in human fibroblasts. Biochem Biophys Res Commun. 1982 Dec 15;109(3):762–768. doi: 10.1016/0006-291x(82)92005-8. [DOI] [PubMed] [Google Scholar]
  13. Owen N. E., Villereal M. L. Evidence for a role of calmodulin in serum stimulation of Na+ influx in human fibroblasts. Proc Natl Acad Sci U S A. 1982 Jun;79(11):3537–3541. doi: 10.1073/pnas.79.11.3537. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Pouysségur J., Chambard J. C., Franchi A., Paris S., Van Obberghen-Schilling E. Growth factor activation of an amiloride-sensitive Na+/H+ exchange system in quiescent fibroblasts: coupling to ribosomal protein S6 phosphorylation. Proc Natl Acad Sci U S A. 1982 Jul;79(13):3935–3939. doi: 10.1073/pnas.79.13.3935. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Pozzan T., Gatti G., Dozio N., Vicentini L. M., Meldolesi J. Ca2+-dependent and -independent release of neurotransmitters from PC12 cells: a role for protein kinase C activation? J Cell Biol. 1984 Aug;99(2):628–638. doi: 10.1083/jcb.99.2.628. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Smith J. B., Rozengurt E. Serum stimulates the Na+,K+ pump in quiescent fibroblasts by increasing Na+ entry. Proc Natl Acad Sci U S A. 1978 Nov;75(11):5560–5564. doi: 10.1073/pnas.75.11.5560. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Takayama S., White M. F., Lauris V., Kahn C. R. Phorbol esters modulate insulin receptor phosphorylation and insulin action in cultured hepatoma cells. Proc Natl Acad Sci U S A. 1984 Dec;81(24):7797–7801. doi: 10.1073/pnas.81.24.7797. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Vicentini L. M., Di Virgilio F., Ambrosini A., Pozzan T., Meldolesi J. Tumor promoter phorbol 12-myristate, 13-acetate inhibits phosphoinositide hydrolysis and cytosolic Ca2+ rise induced by the activation of muscarinic receptors in PC12 cells. Biochem Biophys Res Commun. 1985 Feb 28;127(1):310–317. doi: 10.1016/s0006-291x(85)80160-1. [DOI] [PubMed] [Google Scholar]
  19. Vicentini L. M., Villereal M. L. Serum, bradykinin and vasopressin stimulate release of inositol phosphates from human fibroblasts. Biochem Biophys Res Commun. 1984 Sep 17;123(2):663–670. doi: 10.1016/0006-291x(84)90280-8. [DOI] [PubMed] [Google Scholar]
  20. Villereal M. L. Sodium fluxes in human fibroblasts: effect of serum, Ca+2, and amiloride. J Cell Physiol. 1981 Jun;107(3):359–369. doi: 10.1002/jcp.1041070307. [DOI] [PubMed] [Google Scholar]
  21. Whiteley B., Cassel D., Zhuang Y. X., Glaser L. Tumor promoter phorbol 12-myristate 13-acetate inhibits mitogen-stimulated Na+/H+ exchange in human epidermoid carcinoma A431 cells. J Cell Biol. 1984 Sep;99(3):1162–1166. doi: 10.1083/jcb.99.3.1162. [DOI] [PMC free article] [PubMed] [Google Scholar]

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