Skip to main content
NCBI home page
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1993 Jul;92(1):169–178. doi: 10.1172/JCI116545

Attenuated prostaglandin formation in peroxisomal-deficient human skin fibroblasts.

J A Gordon 1, L J Warnock 1, A A Spector 1
PMCID: PMC293558  PMID: 7686919

Abstract

Peroxisomal-deficient skin fibroblasts from patients with Zellweger's syndrome or infantile Refsum's disease produced fewer prostaglandins than normal skin fibroblasts. Radioimmunoassay indicated a 45-55% decrease in prostaglandin E2 (PGE2) production when Zellweger's fibroblasts were incubated with arachidonic acid. This deficiency was not overcome by pretreatment of the Zellweger's fibroblasts with media containing arachidonic acid, and it was not due to channeling of arachidonic acid into other eicosanoid products. Modifications in the peroxide tone of the Zellweger's fibroblasts by addition of H2O2 or catalase failed to increase PGE2 production. Using Northern analysis, we were unable to detect an mRNA transcript for PGH synthase in unstimulated Zellweger fibroblasts but identified a 4.2-kb mRNA transcript after treatment with phorbol myristate acetate (PMA). Treatment for 6 h with 10 nM PMA raised PGE2 production in normal and Zellweger fibroblasts to equivalent levels. These increases were prevented by addition of H-7, staurosporine, cycloheximide, or actinomycin D. Our findings suggest that the reduced PGE2 production in peroxisomal deficient fibroblasts is due to a decrease in PGH synthase mRNA. The reduction in PGH synthase can be overcome by treatment of the cells with agents which enhance gene expression.

Full text

PDF
169

Images in this article

171Image
on p.171
175Image
on p.175
175Image
on p.175
176Image
on p.176

Selected References

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

  1. Beaudry G. A., Waite M., Daniel L. W. Regulation of arachidonic acid metabolism in Madin-Darby canine kidney cells: stimulation of synthesis of the cyclooxygenase system by 12-O-tetradecanoyl-phorbol-13-acetate. Arch Biochem Biophys. 1985 May 15;239(1):242–247. doi: 10.1016/0003-9861(85)90832-x. [DOI] [PubMed] [Google Scholar]
  2. Beckman J. S., Minor R. L., Jr, White C. W., Repine J. E., Rosen G. M., Freeman B. A. Superoxide dismutase and catalase conjugated to polyethylene glycol increases endothelial enzyme activity and oxidant resistance. J Biol Chem. 1988 May 15;263(14):6884–6892. [PubMed] [Google Scholar]
  3. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  4. Daniel L. W., King L., Waite M. Source of arachidonic acid for prostaglandin synthesis in Madin-Darby canine kidney cells. J Biol Chem. 1981 Dec 25;256(24):12830–12835. [PubMed] [Google Scholar]
  5. De Duve C., Baudhuin P. Peroxisomes (microbodies and related particles). Physiol Rev. 1966 Apr;46(2):323–357. doi: 10.1152/physrev.1966.46.2.323. [DOI] [PubMed] [Google Scholar]
  6. DeWitt D. L. Prostaglandin endoperoxide synthase: regulation of enzyme expression. Biochim Biophys Acta. 1991 May 8;1083(2):121–134. doi: 10.1016/0005-2760(91)90032-d. [DOI] [PubMed] [Google Scholar]
  7. Diaz A., Reginato A. M., Jimenez S. A. Alternative splicing of human prostaglandin G/H synthase mRNA and evidence of differential regulation of the resulting transcripts by transforming growth factor beta 1, interleukin 1 beta, and tumor necrosis factor alpha. J Biol Chem. 1992 May 25;267(15):10816–10822. [PubMed] [Google Scholar]
  8. Diczfalusy U., Alexson S. E. Identification of metabolites from peroxisomal beta-oxidation of prostaglandins. J Lipid Res. 1990 Feb;31(2):307–314. [PubMed] [Google Scholar]
  9. Duniec Z. M., Nettesheim P., Eling T. E. Stimulation of prostaglandin H synthase mRNA levels and prostaglandin biosynthesis by phorbol ester: mediation by protein kinase C. Mol Pharmacol. 1991 Feb;39(2):164–170. [PubMed] [Google Scholar]
  10. Funk C. D., Furci L., FitzGerald G. A. Molecular cloning, primary structure, and expression of the human platelet/erythroleukemia cell 12-lipoxygenase. Proc Natl Acad Sci U S A. 1990 Aug;87(15):5638–5642. doi: 10.1073/pnas.87.15.5638. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Glisin V., Crkvenjakov R., Byus C. Ribonucleic acid isolated by cesium chloride centrifugation. Biochemistry. 1974 Jun 4;13(12):2633–2637. doi: 10.1021/bi00709a025. [DOI] [PubMed] [Google Scholar]
  12. Goldfischer S., Moore C. L., Johnson A. B., Spiro A. J., Valsamis M. P., Wisniewski H. K., Ritch R. H., Norton W. T., Rapin I., Gartner L. M. Peroxisomal and mitochondrial defects in the cerebro-hepato-renal syndrome. Science. 1973 Oct 5;182(4107):62–64. doi: 10.1126/science.182.4107.62. [DOI] [PubMed] [Google Scholar]
  13. Gordon J. A., Figard P. H., Spector A. A. Hydroxyeicosatetraenoic acid metabolism in cultured human skin fibroblasts. Evidence for peroxisomal beta-oxidation. J Clin Invest. 1990 Apr;85(4):1173–1181. doi: 10.1172/JCI114550. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Gordon J. A., Spector A. A. Effects of 12-HETE on renal tubular epithelial cells. Am J Physiol. 1987 Aug;253(2 Pt 1):C277–C285. doi: 10.1152/ajpcell.1987.253.2.C277. [DOI] [PubMed] [Google Scholar]
  15. Habenicht A. J., Goerig M., Grulich J., Rothe D., Gronwald R., Loth U., Schettler G., Kommerell B., Ross R. Human platelet-derived growth factor stimulates prostaglandin synthesis by activation and by rapid de novo synthesis of cyclooxygenase. J Clin Invest. 1985 Apr;75(4):1381–1387. doi: 10.1172/JCI111839. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hajra A. K., Burke C. L., Jones C. L. Subcellular localization of acyl coenzyme A: dihydroxyacetone phosphate acyltransferase in rat liver peroxisomes (microbodies). J Biol Chem. 1979 Nov 10;254(21):10896–10900. [PubMed] [Google Scholar]
  17. Han J. W., Sadowski H., Young D. A., Macara I. G. Persistent induction of cyclooxygenase in p60v-src-transformed 3T3 fibroblasts. Proc Natl Acad Sci U S A. 1990 May;87(9):3373–3377. doi: 10.1073/pnas.87.9.3373. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hemler M. E., Cook H. W., Lands W. E. Prostaglandin biosynthesis can be triggered by lipid peroxides. Arch Biochem Biophys. 1979 Apr 1;193(2):340–345. doi: 10.1016/0003-9861(79)90038-9. [DOI] [PubMed] [Google Scholar]
  19. Hiltunen J. K., Kärki T., Hassinen I. E., Osmundsen H. beta-Oxidation of polyunsaturated fatty acids by rat liver peroxisomes. A role for 2,4-dienoyl-coenzyme A reductase in peroxisomal beta-oxidation. J Biol Chem. 1986 Dec 15;261(35):16484–16493. [PubMed] [Google Scholar]
  20. Hla T., Neilson K. Human cyclooxygenase-2 cDNA. Proc Natl Acad Sci U S A. 1992 Aug 15;89(16):7384–7388. doi: 10.1073/pnas.89.16.7384. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kelley R. I. Review: the cerebrohepatorenal syndrome of Zellweger, morphologic and metabolic aspects. Am J Med Genet. 1983 Dec;16(4):503–517. doi: 10.1002/ajmg.1320160409. [DOI] [PubMed] [Google Scholar]
  22. Krisans S. K., Thompson S. L., Pena L. A., Kok E., Javitt N. B. Bile acid synthesis in rat liver peroxisomes: metabolism of 26-hydroxycholesterol to 3 beta-hydroxy-5-cholenoic acid. J Lipid Res. 1985 Nov;26(11):1324–1332. [PubMed] [Google Scholar]
  23. Kujubu D. A., Fletcher B. S., Varnum B. C., Lim R. W., Herschman H. R. TIS10, a phorbol ester tumor promoter-inducible mRNA from Swiss 3T3 cells, encodes a novel prostaglandin synthase/cyclooxygenase homologue. J Biol Chem. 1991 Jul 15;266(20):12866–12872. [PubMed] [Google Scholar]
  24. Kujubu D. A., Herschman H. R. Dexamethasone inhibits mitogen induction of the TIS10 prostaglandin synthase/cyclooxygenase gene. J Biol Chem. 1992 Apr 25;267(12):7991–7994. [PubMed] [Google Scholar]
  25. Labarca C., Paigen K. A simple, rapid, and sensitive DNA assay procedure. Anal Biochem. 1980 Mar 1;102(2):344–352. doi: 10.1016/0003-2697(80)90165-7. [DOI] [PubMed] [Google Scholar]
  26. 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]
  27. Lazarow P. B. Rat liver peroxisomes catalyze the beta oxidation of fatty acids. J Biol Chem. 1978 Mar 10;253(5):1522–1528. [PubMed] [Google Scholar]
  28. Lees M. B., Paxman S. Modification of the Lowry procedure for the analysis of proteolipid protein. Anal Biochem. 1972 May;47(1):184–192. doi: 10.1016/0003-2697(72)90291-6. [DOI] [PubMed] [Google Scholar]
  29. Maier J. A., Hla T., Maciag T. Cyclooxygenase is an immediate-early gene induced by interleukin-1 in human endothelial cells. J Biol Chem. 1990 Jul 5;265(19):10805–10808. [PubMed] [Google Scholar]
  30. Marshall P. J., Kulmacz R. J., Lands W. E. Constraints on prostaglandin biosynthesis in tissues. J Biol Chem. 1987 Mar 15;262(8):3510–3517. [PubMed] [Google Scholar]
  31. O'Banion M. K., Winn V. D., Young D. A. cDNA cloning and functional activity of a glucocorticoid-regulated inflammatory cyclooxygenase. Proc Natl Acad Sci U S A. 1992 Jun 1;89(11):4888–4892. doi: 10.1073/pnas.89.11.4888. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Pagels W. R., Sachs R. J., Marnett L. J., Dewitt D. L., Day J. S., Smith W. L. Immunochemical evidence for the involvement of prostaglandin H synthase in hydroperoxide-dependent oxidations by ram seminal vesicle microsomes. J Biol Chem. 1983 May 25;258(10):6517–6523. [PubMed] [Google Scholar]
  33. Parker J., Daniel L. W., Waite M. Evidence of protein kinase C involvement in phorbol diester-stimulated arachidonic acid release and prostaglandin synthesis. J Biol Chem. 1987 Apr 15;262(11):5385–5393. [PubMed] [Google Scholar]
  34. Poulos A., Sharp P., Whiting M. Infantile Refsum's disease (phytanic acid storage disease): a variant of Zellweger's syndrome? Clin Genet. 1984 Dec;26(6):579–586. doi: 10.1111/j.1399-0004.1984.tb01107.x. [DOI] [PubMed] [Google Scholar]
  35. Rollins T. E., Smith W. L. Subcellular localization of prostaglandin-forming cyclooxygenase in Swiss mouse 3T3 fibroblasts by electron microscopic immunocytochemistry. J Biol Chem. 1980 May 25;255(10):4872–4875. [PubMed] [Google Scholar]
  36. Schrakamp G., Schalkwijk C. G., Schutgens R. B., Wanders R. J., Tager J. M., van den Bosch H. Plasmalogen biosynthesis in peroxisomal disorders: fatty alcohol versus alkylglycerol precursors. J Lipid Res. 1988 Mar;29(3):325–334. [PubMed] [Google Scholar]
  37. Tiffany C. W., Hoefler S., Moser H. W., Burch R. M. Arachidonic acid metabolism in fibroblasts from patients with peroxisomal diseases: response to interleukin 1. Biochim Biophys Acta. 1990 Nov 14;1096(1):41–46. doi: 10.1016/0925-4439(90)90010-m. [DOI] [PubMed] [Google Scholar]
  38. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Trijbels J. M., Monnens L. A., Bakkeren J. A., Van Raay-Selten A. H., Corstiaensen J. M. Biochemical studies in the cerebro-hepato-renal syndrome of Zellweger: a disturbance in the metabolism of pipecolic acid. J Inherit Metab Dis. 1980;2(2):39–42. doi: 10.1007/BF01799073. [DOI] [PubMed] [Google Scholar]
  40. Wanders R. J., Kos M., Roest B., Meijer A. J., Schrakamp G., Heymans H. S., Tegelaers W. H., van den Bosch H., Schutgens R. B., Tager J. M. Activity of peroxisomal enzymes and intracellular distribution of catalase in Zellweger syndrome. Biochem Biophys Res Commun. 1984 Sep 28;123(3):1054–1061. doi: 10.1016/s0006-291x(84)80240-5. [DOI] [PubMed] [Google Scholar]
  41. Whorton A. R., Montgomery M. E., Kent R. S. Effect of hydrogen peroxide on prostaglandin production and cellular integrity in cultured porcine aortic endothelial cells. J Clin Invest. 1985 Jul;76(1):295–302. doi: 10.1172/JCI111960. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Wu K. K., Hatzakis H., Lo S. S., Seong D. C., Sanduja S. K., Tai H. H. Stimulation of de novo synthesis of prostaglandin G/H synthase in human endothelial cells by phorbol ester. J Biol Chem. 1988 Dec 15;263(35):19043–19047. [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES