Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 2000 Mar 15;346(Pt 3):751–758.

Involvement of lipoxygenase in lysophosphatidic acid-stimulated hydrogen peroxide release in human HaCaT keratinocytes.

M Sekharam 1, J M Cunnick 1, J Wu 1
PMCID: PMC1220909  PMID: 10698703

Abstract

Although it is now recognized that low levels of reactive oxygen species (ROS) are required for the mitogenic response, mitogen-induced signalling pathways that regulate ROS generation in non-phagocytic cells remain largely uncharacterized. Using a real-time assay for measuring hydrogen peroxide (H(2)O(2)) formation, we analysed H(2)O(2) release in human HaCaT keratinocytes in response to lysophosphatidic acid (LPA), a mitogen for keratinocytes. LPA rapidly increased H(2)O(2) release in HaCaT cells. Unlike LPA-induced mitogen-activated protein (MAP) kinase activation, LPA-stimulated H(2)O(2) release was independent of the tyrosine kinase activity of the epidermal growth factor (EGF) receptor. Calcium chelators, phospholipase A(2) inhibitors, and lipoxygenase inhibitors effectively blocked LPA-stimulated H(2)O(2) release, whereas cyclooxygenase inhibitors were without effect. Addition of 5-lipoxygenase products 5-hydroperoxyeicosatetraenoic acid (5-HPETE) and leukotriene B(4), but not 5-hydroxyeicosatetraenoic acid (5-HETE) and leukotriene C(4), restored LPA-stimulated H(2)O(2) release in cells treated with the lipoxygenase inhibitors nordihydroguaiaretic acid and Zileuton. These results suggest that the lipoxygenase products 5-HPETE and leukotriene B(4) are required for LPA-stimulated H(2)O(2) release in HaCaT cells.

Full Text

The Full Text of this article is available as a PDF (285.1 KB).

Selected References

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

  1. An S., Bleu T., Zheng Y., Goetzl E. J. Recombinant human G protein-coupled lysophosphatidic acid receptors mediate intracellular calcium mobilization. Mol Pharmacol. 1998 Nov;54(5):881–888. doi: 10.1124/mol.54.5.881. [DOI] [PubMed] [Google Scholar]
  2. Aoki Y., Qiu D., Zhao G. H., Kao P. N. Leukotriene B4 mediates histamine induction of NF-kappaB and IL-8 in human bronchial epithelial cells. Am J Physiol. 1998 Jun;274(6 Pt 1):L1030–L1039. doi: 10.1152/ajplung.1998.274.6.L1030. [DOI] [PubMed] [Google Scholar]
  3. Bae Y. S., Kang S. W., Seo M. S., Baines I. C., Tekle E., Chock P. B., Rhee S. G. Epidermal growth factor (EGF)-induced generation of hydrogen peroxide. Role in EGF receptor-mediated tyrosine phosphorylation. J Biol Chem. 1997 Jan 3;272(1):217–221. [PubMed] [Google Scholar]
  4. Bonizzi G., Piette J., Schoonbroodt S., Greimers R., Havard L., Merville M. P., Bours V. Reactive oxygen intermediate-dependent NF-kappaB activation by interleukin-1beta requires 5-lipoxygenase or NADPH oxidase activity. Mol Cell Biol. 1999 Mar;19(3):1950–1960. doi: 10.1128/mcb.19.3.1950. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Carter G. W., Young P. R., Albert D. H., Bouska J., Dyer R., Bell R. L., Summers J. B., Brooks D. W. 5-lipoxygenase inhibitory activity of zileuton. J Pharmacol Exp Ther. 1991 Mar;256(3):929–937. [PubMed] [Google Scholar]
  6. Chen Q., Olashaw N., Wu J. Participation of reactive oxygen species in the lysophosphatidic acid-stimulated mitogen-activated protein kinase kinase activation pathway. J Biol Chem. 1995 Dec 1;270(48):28499–28502. doi: 10.1074/jbc.270.48.28499. [DOI] [PubMed] [Google Scholar]
  7. Cunnick J. M., Dorsey J. F., Standley T., Turkson J., Kraker A. J., Fry D. W., Jove R., Wu J. Role of tyrosine kinase activity of epidermal growth factor receptor in the lysophosphatidic acid-stimulated mitogen-activated protein kinase pathway. J Biol Chem. 1998 Jun 5;273(23):14468–14475. doi: 10.1074/jbc.273.23.14468. [DOI] [PubMed] [Google Scholar]
  8. Daub H., Wallasch C., Lankenau A., Herrlich A., Ullrich A. Signal characteristics of G protein-transactivated EGF receptor. EMBO J. 1997 Dec 1;16(23):7032–7044. doi: 10.1093/emboj/16.23.7032. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Daub H., Weiss F. U., Wallasch C., Ullrich A. Role of transactivation of the EGF receptor in signalling by G-protein-coupled receptors. Nature. 1996 Feb 8;379(6565):557–560. doi: 10.1038/379557a0. [DOI] [PubMed] [Google Scholar]
  10. Glaser K. B., Mobilio D., Chang J. Y., Senko N. Phospholipase A2 enzymes: regulation and inhibition. Trends Pharmacol Sci. 1993 Mar;14(3):92–98. doi: 10.1016/0165-6147(93)90071-q. [DOI] [PubMed] [Google Scholar]
  11. Goetzl E. J., An S. Diversity of cellular receptors and functions for the lysophospholipid growth factors lysophosphatidic acid and sphingosine 1-phosphate. FASEB J. 1998 Dec;12(15):1589–1598. [PubMed] [Google Scholar]
  12. Goldman R., Moshonov S., Zor U. Generation of reactive oxygen species in a human keratinocyte cell line: role of calcium. Arch Biochem Biophys. 1998 Feb 1;350(1):10–18. doi: 10.1006/abbi.1997.0478. [DOI] [PubMed] [Google Scholar]
  13. Halliwell B., Gutteridge J. M. Role of free radicals and catalytic metal ions in human disease: an overview. Methods Enzymol. 1990;186:1–85. doi: 10.1016/0076-6879(90)86093-b. [DOI] [PubMed] [Google Scholar]
  14. Irani K., Xia Y., Zweier J. L., Sollott S. J., Der C. J., Fearon E. R., Sundaresan M., Finkel T., Goldschmidt-Clermont P. J. Mitogenic signaling mediated by oxidants in Ras-transformed fibroblasts. Science. 1997 Mar 14;275(5306):1649–1652. doi: 10.1126/science.275.5306.1649. [DOI] [PubMed] [Google Scholar]
  15. Jalink K., Hengeveld T., Mulder S., Postma F. R., Simon M. F., Chap H., van der Marel G. A., van Boom J. H., van Blitterswijk W. J., Moolenaar W. H. Lysophosphatidic acid-induced Ca2+ mobilization in human A431 cells: structure-activity analysis. Biochem J. 1995 Apr 15;307(Pt 2):609–616. doi: 10.1042/bj3070609. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Janssen-Timmen U., Vickers P. J., Wittig U., Lehmann W. D., Stark H. J., Fusenig N. E., Rosenbach T., Rådmark O., Samuelsson B., Habenicht A. J. Expression of 5-lipoxygenase in differentiating human skin keratinocytes. Proc Natl Acad Sci U S A. 1995 Jul 18;92(15):6966–6970. doi: 10.1073/pnas.92.15.6966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Joneson T., Bar-Sagi D. A Rac1 effector site controlling mitogenesis through superoxide production. J Biol Chem. 1998 Jul 17;273(29):17991–17994. doi: 10.1074/jbc.273.29.17991. [DOI] [PubMed] [Google Scholar]
  18. Klohs W. D., Fry D. W., Kraker A. J. Inhibitors of tyrosine kinase. Curr Opin Oncol. 1997 Nov;9(6):562–568. doi: 10.1097/00001622-199711000-00012. [DOI] [PubMed] [Google Scholar]
  19. Lanni C., Becker E. L. Inhibition of neutrophil phospholipase A2 by p-bromophenylacyl bromide, nordihydroguaiaretic acid, 5,8,11,14-eicosatetraynoic acid and quercetin. Int Arch Allergy Appl Immunol. 1985;76(3):214–217. doi: 10.1159/000233694. [DOI] [PubMed] [Google Scholar]
  20. Lo Y. Y., Cruz T. F. Involvement of reactive oxygen species in cytokine and growth factor induction of c-fos expression in chondrocytes. J Biol Chem. 1995 May 19;270(20):11727–11730. doi: 10.1074/jbc.270.20.11727. [DOI] [PubMed] [Google Scholar]
  21. Los M., Schenk H., Hexel K., Baeuerle P. A., Dröge W., Schulze-Osthoff K. IL-2 gene expression and NF-kappa B activation through CD28 requires reactive oxygen production by 5-lipoxygenase. EMBO J. 1995 Aug 1;14(15):3731–3740. doi: 10.1002/j.1460-2075.1995.tb00043.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Luttrell L. M., Della Rocca G. J., van Biesen T., Luttrell D. K., Lefkowitz R. J. Gbetagamma subunits mediate Src-dependent phosphorylation of the epidermal growth factor receptor. A scaffold for G protein-coupled receptor-mediated Ras activation. J Biol Chem. 1997 Feb 14;272(7):4637–4644. doi: 10.1074/jbc.272.7.4637. [DOI] [PubMed] [Google Scholar]
  23. Moolenaar W. H., Kranenburg O., Postma F. R., Zondag G. C. Lysophosphatidic acid: G-protein signalling and cellular responses. Curr Opin Cell Biol. 1997 Apr;9(2):168–173. doi: 10.1016/s0955-0674(97)80059-2. [DOI] [PubMed] [Google Scholar]
  24. Moolenaar W. H. Lysophosphatidic acid, a multifunctional phospholipid messenger. J Biol Chem. 1995 Jun 2;270(22):12949–12952. doi: 10.1074/jbc.270.22.12949. [DOI] [PubMed] [Google Scholar]
  25. Rewcastle G. W., Palmer B. D., Thompson A. M., Bridges A. J., Cody D. R., Zhou H., Fry D. W., McMichael A., Denny W. A. Tyrosine kinase inhibitors. 10. Isomeric 4-[(3-bromophenyl)amino]pyrido[d]-pyrimidines are potent ATP binding site inhibitors of the tyrosine kinase function of the epidermal growth factor receptor. J Med Chem. 1996 Apr 26;39(9):1823–1835. doi: 10.1021/jm9508651. [DOI] [PubMed] [Google Scholar]
  26. Rosenthal M. D., Vishwanath B. S., Franson R. C. Effects of aristolochic acid on phospholipase A2 activity and arachidonate metabolism of human neutrophils. Biochim Biophys Acta. 1989 Jan 23;1001(1):1–8. doi: 10.1016/0005-2760(89)90299-3. [DOI] [PubMed] [Google Scholar]
  27. Sekharam M., Trotti A., Cunnick J. M., Wu J. Suppression of fibroblast cell cycle progression in G1 phase by N-acetylcysteine. Toxicol Appl Pharmacol. 1998 Apr;149(2):210–216. doi: 10.1006/taap.1997.8361. [DOI] [PubMed] [Google Scholar]
  28. Sundaresan M., Yu Z. X., Ferrans V. J., Irani K., Finkel T. Requirement for generation of H2O2 for platelet-derived growth factor signal transduction. Science. 1995 Oct 13;270(5234):296–299. doi: 10.1126/science.270.5234.296. [DOI] [PubMed] [Google Scholar]
  29. Sundaresan M., Yu Z. X., Ferrans V. J., Sulciner D. J., Gutkind J. S., Irani K., Goldschmidt-Clermont P. J., Finkel T. Regulation of reactive-oxygen-species generation in fibroblasts by Rac1. Biochem J. 1996 Sep 1;318(Pt 2):379–382. doi: 10.1042/bj3180379. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. van Corven E. J., Groenink A., Jalink K., Eichholtz T., Moolenaar W. H. Lysophosphatidate-induced cell proliferation: identification and dissection of signaling pathways mediated by G proteins. Cell. 1989 Oct 6;59(1):45–54. doi: 10.1016/0092-8674(89)90868-4. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES