Journal of Molecular Medicine

, Volume 87, Issue 6, pp 645–657

Involvement of the ABC-transporter ABCC1 and the sphingosine 1-phosphate receptor subtype S1P3 in the cytoprotection of human fibroblasts by the glucocorticoid dexamethasone

  • Barbara Nieuwenhuis
  • Anja Lüth
  • Jerold Chun
  • Andrea Huwiler
  • Josef Pfeilschifter
  • Monika Schäfer-Korting
  • Burkhard Kleuser
Original Article


Glucocorticoids (GC) represent the most commonly used drugs for the treatment of acute and chronic inflammatory skin diseases. However, the topical long-term therapy of GC is limited by the occurrence of skin atrophy. Most interestingly, although GC inhibit proliferation of human fibroblasts, they exert a pronounced anti-apoptopic action. In the present study, we further elucidated the molecular mechanism of the GC dexamethasone (Dex) to protect human fibroblasts from programmed cell death. Dex not only significantly alters the expression of the cytosolic isoenzyme sphingosine kinase 1 but also initiated an enhanced intracellular formation of the sphingolipid sphingosine 1-phosphate (S1P). Investigations using S1P3(−/−)-fibroblasts revealed that this S1P-receptor subtype is essential for the Dex-induced cytoprotection. Moreover, we demonstrate that the ATP-binding cassette (ABC)-transporter ABCC1 is upregulated by Dex and may represent a crucial carrier to transport S1P from the cytosol to the S1P3-receptor subtype.


Glucocorticoids Dexamethasone Sphingosine kinase Sphingosine 1-phosphate ABC-transporter S1P-receptor subtypes 



ATP-binding cassette transporter


actinomycin D




Bcl-2 homology 3 domain


bovine serum albumin


Dulbecco’s modified Eagle’s medium


ethylenediaminetetraacetic acid


epidermal growth factor


fluorescence-activated cell sorting


foetal calf serum


FADD-like IL-1β-converting enzyme inhibitory protein




FADD-like IL-1β-converting enzyme




G-protein-coupled receptor


glucocorticoid receptor


4-(2-Hydroxyethyl)-1-piperazineethanesulfonic acid




mapkinase phosphatase 1


nuclear factor k B




phosphate-buffered saline


polymerase chain reaction


platelet-derived growth factor




polyvinylidene difluoride


sodium dodecyl sulphate


small interfering RNA


sphingosine kinase


sphingosine 1-phosphate


tumour necrosis factor α


vascular endothelial growth factor

Supplementary material

109_2009_468_MOESM1_ESM.pdf (497 kb)
ESM 1(PDF 497 kb)


  1. 1.
    Rhen T, Cidlowski JA (2005) Antiinflammatory action of glucocorticoids new mechanisms for old drugs. N Engl J Med 353:1711–1723PubMedCrossRefGoogle Scholar
  2. 2.
    Schoepe S, Schacke H, May E et al (2006) Glucocorticoid therapy-induced skin atrophy. Exp Dermatol 15:406–420PubMedCrossRefGoogle Scholar
  3. 3.
    Hammer S, Sauer B, Spika I et al (2004) Glucocorticoids mediate differential anti-apoptotic effects in human fibroblasts and keratinocytes via sphingosine-1-phosphate formation. J Cell Biochem 91:840–851PubMedCrossRefGoogle Scholar
  4. 4.
    Sharma S, Lichtenstein A (2008) Dexamethasone-induced apoptotic mechanisms in myeloma cells investigated by analysis of mutant glucocorticoid receptors. Blood 112:1338–1345PubMedCrossRefGoogle Scholar
  5. 5.
    Herr I, Gassler N, Friess H et al (2007) Regulation of differential pro- and anti-apoptotic signaling by glucocorticoids. Apoptosis 12:271–291PubMedCrossRefGoogle Scholar
  6. 6.
    Oh HY, Namkoong S, Lee SJ et al (2006) Dexamethasone protects primary cultured hepatocytes from death receptor-mediated apoptosis by upregulation of cFLIP. Cell Death Differ 13:512–523PubMedCrossRefGoogle Scholar
  7. 7.
    Spiegel S, Milstien S (2002) Sphingosine 1-phosphate, a key cell signaling molecule. J Biol Chem 277:25851–25854PubMedCrossRefGoogle Scholar
  8. 8.
    Takabe K, Paugh SW, Milstien S et al (2008) "Inside-out" signaling of sphingosine-1-phosphate: therapeutic targets. Pharmacol Rev 60:181–195PubMedCrossRefGoogle Scholar
  9. 9.
    Watterson KR, Lanning DA, Diegelmann RF et al (2007) Regulation of fibroblast functions by lysophospholipid mediators: potential roles in wound healing. Wound Repair Regen 15:607–616PubMedCrossRefGoogle Scholar
  10. 10.
    Hofmann LP, Ren S, Schwalm S et al (2008) Sphingosine kinase 1 and 2 regulate the capacity of mesangial cells to resist apoptotic stimuli in an opposing manner. Biol Chem 389:1399–1407PubMedCrossRefGoogle Scholar
  11. 11.
    Sukocheva OA, Wang L, Albanese N et al (2003) Sphingosine kinase transmits estrogen signaling in human breast cancer cells. Mol Endocrinol 17:2002–2012PubMedCrossRefGoogle Scholar
  12. 12.
    Goetzl EJ, Wang W, McGiffert C et al (2007) Sphingosine 1-phosphate as an intracellular messenger and extracellular mediator in immunity. Acta Paediatr Suppl 96:49–52PubMedCrossRefGoogle Scholar
  13. 13.
    Keller CD, Rivera Gil P, Tolle M et al (2007) Immunomodulator FTY720 induces myofibroblast differentiation via the lysophospholipid receptor S1P3 and Smad3 signaling. Am J Pathol 170:281–292PubMedCrossRefGoogle Scholar
  14. 14.
    Mitra P, Oskeritzian CA, Payne SG et al (2006) Role of ABCC1 in export of sphingosine-1-phosphate from mast cells. Proc Natl Acad Sci USA 103:16394–16399PubMedCrossRefGoogle Scholar
  15. 15.
    Sato K, Malchinkhuu E, Horiuchi Y et al (2007) Critical role of ABCA1 transporter in sphingosine 1-phosphate release from astrocytes. J Neurochem 103:2610–2619Google Scholar
  16. 16.
    Ishii I, Friedman B, Ye X et al (2001) Selective loss of sphingosine 1-phosphate signaling with no obvious phenotypic abnormality in mice lacking its G protein-coupled receptor, LP(B3)/EDG-3. J Biol Chem 276:33697–33704PubMedCrossRefGoogle Scholar
  17. 17.
    Manggau M, Kim DS, Ruwisch L et al (2001) 1Alpha, 25-Dihydroxyvitamnin D3 protects human keratinocytes from apoptosis by the formation of sphingosine 1-phosphate. J Invest Dermatol 117:1241–1249PubMedCrossRefGoogle Scholar
  18. 18.
    Ruwisch L, Schafer-Korting M, Kleuser B (2001) An improved high-performance liquid chromatographic method for the determination of sphingosine-1-phosphate in complex biological materials. Naunyn Schmiedeberg’s Arch Pharmacol 363:358–363CrossRefGoogle Scholar
  19. 19.
    Ayaori M, Sawada S, Yonemura A et al (2006) Glucocorticoid receptor regulates ATP-binding cassette transporter-A1 expression and apolipoprotein-mediated cholesterol efflux from macrophages. Arterioscler Thromb Vasc Biol 26:163–168PubMedCrossRefGoogle Scholar
  20. 20.
    Amsterdam A, Tajima K, Sasson R (2002) Cell-specific regulation of apoptosis by glucocorticoids: implication to their anti-inflammatory action. Biochem Pharmacol 64:843–850PubMedCrossRefGoogle Scholar
  21. 21.
    Schmidt M, Pauels HG, Lugering N et al (1999) Glucocorticoids induce apoptosis in human monocytes: potential role of IL-1 beta. J Immunol 163:3484–3490PubMedGoogle Scholar
  22. 22.
    Davis MC, McColl KS, Zhong F et al (2008) Dexamethasone-induced inositol 1, 4, 5-trisphosphate receptor elevation in murine lymphoma cells is not required for dexamethasone-mediated calcium elevation and apoptosis. J Biol Chem 283:10357–10365PubMedCrossRefGoogle Scholar
  23. 23.
    Tissing WJ, Meijerink JP, den Boer ML et al (2003) Molecular determinants of glucocorticoid sensitivity and resistance in acute lymphoblastic leukemia. Leukemia 17:17–25PubMedCrossRefGoogle Scholar
  24. 24.
    Chittenden T (2002) BH3 domains: intracellular death-ligands critical for initiating apoptosis. Cancer Cell 2:165–166PubMedCrossRefGoogle Scholar
  25. 25.
    Liu H, Toman RE, Goparaju SK et al (2003) Sphingosine kinase type 2 is a putative BH3-only protein that induces apoptosis. J Biol Chem 278:40330–40336PubMedCrossRefGoogle Scholar
  26. 26.
    Letai A, Bassik MC, Walensky LD et al (2002) Distinct BH3 domains either sensitize or activate mitochondrial apoptosis, serving as prototype cancer therapeutics. Cancer Cell 2:183–192PubMedCrossRefGoogle Scholar
  27. 27.
    Saffar AS, Dragon S, Ezzati P et al (2008) Phosphatidylinositol 3-kinase and p38 mitogen-activated protein kinase regulate induction of Mcl-1 and survival in glucocorticoid-treated human neutrophils. J Allergy Clin Immunol 121:492–498PubMedCrossRefGoogle Scholar
  28. 28.
    Evans-Storms RB, Cidlowski JA (2000) Delineation of an antiapoptotic action of glucocorticoids in hepatoma cells: the role of nuclear factor-kappaB. Endocrinol 141:1854–1862CrossRefGoogle Scholar
  29. 29.
    Sauer B, Gonska H, Manggau M et al (2005) Sphingosine 1-phosphate is involved in cytoprotective actions of calcitriol in human fibroblasts and enhances the intracellular Bcl-2/Bax rheostat. Pharmazie 60:298–304PubMedGoogle Scholar
  30. 30.
    Shida D, Takabe K, Kapitonov D et al (2008) Targeting SphK1 as a new strategy against cancer. Curr Drug Targets 9:662–673PubMedCrossRefGoogle Scholar
  31. 31.
    Taha TA, Argraves KM, Obeid LM (2004) Sphingosine-1-phosphate receptors: receptor specificity versus functional redundancy. Biochim Biophys Acta 1682:48–55PubMedGoogle Scholar
  32. 32.
    Sankala HM, Hait NC, Paugh SW et al (2007) Involvement of sphingosine kinase 2 in p53-independent induction of p21 by the chemotherapeutic drug doxorubicin. Cancer Res 67:10466–10474PubMedCrossRefGoogle Scholar
  33. 33.
    Hait NC, Oskeritzian CA, Paugh SW et al (2006) Sphingosine kinases, sphingosine 1-phosphate, apoptosis and diseases. Biochim Biophys Acta 1758:2016–2026PubMedCrossRefGoogle Scholar
  34. 34.
    Kobayashi N, Nishi T, Hirata T et al (2006) Sphingosine 1-phosphate is released from the cytosol of rat platelets in a carrier-mediated manner. J Lipid Res 47:614–621PubMedCrossRefGoogle Scholar
  35. 35.
    Martin P, Riley R, Back DJ et al (2008) Comparison of the induction profile for drug disposition proteins by typical nuclear receptor activators in human hepatic and intestinal cells. Br J Pharmacol 153:805–819PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Barbara Nieuwenhuis
    • 1
  • Anja Lüth
    • 1
  • Jerold Chun
    • 2
  • Andrea Huwiler
    • 3
  • Josef Pfeilschifter
    • 4
  • Monika Schäfer-Korting
    • 1
  • Burkhard Kleuser
    • 1
  1. 1.Institute of Pharmacy, Pharmacology and ToxicologyFreie Universität BerlinBerlinGermany
  2. 2.Department of Molecular Biology, The Scripps Research InstituteLa JollaUSA
  3. 3.Institute of PharmacologyUniversity of BernBernSwitzerland
  4. 4.Pharmazentrum FrankfurtKlinikum der Johann Wolfgang Goethe-UniversitätFrankfurtGermany

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