Molecular Biology Reports

, Volume 38, Issue 2, pp 833–840 | Cite as

All-trans- and 9-cis-retinoic acids activate the human cyclooxynase-2 gene: a role for DR1 as RARE or RXRE

  • Kyuyong Han
  • Irene Moon
  • Hyunjung J. Lim


The human COX-2 promoter contains a direct repeat 1 (DR1) which was shown to confer responsiveness to PPARs. We found that in AN3CA and F9 cells, this hCOX-2 DR1 mediates responsiveness to all-trans-retinoic acid (tRA) or 9-cis-retinoic acid (9cRA), but this effect was suppressed by PPARδ. Truncated PPARδ lacking the activation domain AF2 cannot suppress RA-induced activation of the hCOX-2 gene via DR1, suggesting that cofactor recruitment by AF2 is required for the suppression by PPARδ. Gel shift assay showed that PPAR/RXR, RARβ/RXR, and RXR/RXR, bind to hCOX-2 DR1, revealing the promiscuity of this DR1. Particularly, RXR homodimer was able to bind to this DR1 only in the presence of 9cRA. Our results established that tRA and 9cRA are potent inducers of hCOX-2 and that the hCOX-2 DR1 could either serve as RARE or RXRE depending on cellular contexts.


COX-2 PPARδ RAR RXR DR1 Retinoic acids 





Peroxisome proliferator-activated receptor


Retinoic acid receptor


All-trans-retinoic acid


9-Cis-retinoic acid


Direct repeat 1





This work was supported by National Research Foundation of Korea Grant funded by the Korean Government (313-2008-2-C00642). Authors thank members of the Lim laboratory for various technical supports.


  1. 1.
    Dannenberg AJ, Lippman SM, Mann JR, Subbaramaiah K, DuBois RN (2005) Cyclooxygenase-2 and epidermal growth factor receptor: pharmacologic targets for chemoprevention. J Clin Oncol 23:254–266CrossRefPubMedGoogle Scholar
  2. 2.
    Tsatsanis C, Androulidaki A, Venihaki M, Margioris AN (2006) Signalling networks regulating cyclooxygenase-2. Int J Biochem Cell Biol 38:1654–1661CrossRefPubMedGoogle Scholar
  3. 3.
    Kang YJ, Wingerd BA, Arakawa T, Smith WL (2006) Cyclooxygenase-2 gene transcription in a macrophage model of inflammation. J Immunol 177:8111–8122PubMedGoogle Scholar
  4. 4.
    Zhang X, Zhang J, Yang X, Han X (2007) Several transcription factors regulate COX-2 gene expression in pancreatic beta-cells. Mol Biol Rep 34:199–206CrossRefPubMedGoogle Scholar
  5. 5.
    Meade EA, McIntyre TM, Zimmerman GA, Prescott SM (1999) Peroxisome proliferators enhance cyclooxygenase-2 expression in epithelial cells. J Biol Chem 274:8328–8334CrossRefPubMedGoogle Scholar
  6. 6.
    Kalajdzic T, Faour WH, He QW, Fahmi H, Martel-Pelletier J, Pelletier JP, Di Battista JA (2002) Nimesulide, a preferential cyclooxygenase 2 inhibitor, suppresses peroxisome proliferator-activated receptor induction of cyclooxygenase 2 gene expression in human synovial fibroblasts: evidence for receptor antagonism. Arthritis Rheum 46:494–506CrossRefPubMedGoogle Scholar
  7. 7.
    Mangelsdorf DJ, Evans RM (1995) The RXR heterodimers and orphan receptors. Cell 83:841–850CrossRefPubMedGoogle Scholar
  8. 8.
    Zhang XK, Lehmann J, Hoffmann B, Dawson MI, Cameron J, Graupner G, Hermann T, Tran P, Pfahl M (1992) Homodimer formation of retinoid X receptor induced by 9-cis retinoic acid. Nature 358:587–591CrossRefPubMedGoogle Scholar
  9. 9.
    Nakshatri H, Chambon P (1994) The directly repeated RG(G/T)TCA motifs of the rat and mouse cellular retinol-binding protein II genes are promiscuous binding sites for RAR, RXR, HNF-4, and ARP-1 homo- and heterodimers. J Biol Chem 269:890–902PubMedGoogle Scholar
  10. 10.
    Han K, Song H, Moon I, Augustin R, Moley K, Rogers M, Lim H (2007) Utilization of DR1 as true RARE in regulating the Ssm, a novel retinoic acid-target gene in the mouse testis. J Endocrinol 192:539–551CrossRefPubMedGoogle Scholar
  11. 11.
    Lim H, Gupta RA, Ma WG, Paria BC, Moller DE, Morrow JD, DuBois RN, Trzaskos JM, Dey SK (1999) Cyclo-oxygenase-2-derived prostacyclin mediates embryo implantation in the mouse via PPARdelta. Genes Dev 13:1561–1574CrossRefPubMedGoogle Scholar
  12. 12.
    Lim HJ, Moon I, Han K (2004) Transcriptional cofactors exhibit differential preference toward peroxisome proliferator-activated receptors alpha and delta in uterine cells. Endocrinology 145:2886–2895CrossRefPubMedGoogle Scholar
  13. 13.
    Rochette-Egly C, Chambon P (2001) F9 embryocarcinoma cells: a cell autonomous model to study the functional selectivity of RARs and RXRs in retinoid signaling. Histol Histopathol 16:909–922PubMedGoogle Scholar
  14. 14.
    Gronemeyer H, Gustafsson JA, Laudet V (2004) Principles for modulation of the nuclear receptor superfamily. Nat Rev Drug Discov 3:950–964CrossRefPubMedGoogle Scholar
  15. 15.
    Mader S, Leroy P, Chen JY, Chambon P (1993) Multiple parameters control the selectivity of nuclear receptors for their response elements. Selectivity and promiscuity in response element recognition by retinoic acid receptors and retinoid X receptors. J Biol Chem 268:591–600PubMedGoogle Scholar
  16. 16.
    Chambon P (1996) A decade of molecular biology of retinoic acid receptors. FASEB J 10:940–954PubMedGoogle Scholar
  17. 17.
    Mangelsdorf DJ, Umesono K, Kliewer SA, Borgmeyer U, Ong ES, Evans RM (1991) A direct repeat in the cellular retinol-binding protein type II gene confers differential regulation by RXR and RAR. Cell 66:555–561CrossRefPubMedGoogle Scholar
  18. 18.
    Durand B, Saunders M, Leroy P, Leid M, Chambon P (1992) All-trans and 9-cis retinoic acid induction of CRABPII transcription is mediated by RAR-RXR heterodimers bound to DR1 and DR2 repeated motifs. Cell 71:73–85CrossRefPubMedGoogle Scholar
  19. 19.
    Kurokawa R, DiRenzo J, Boehm M, Sugarman J, Gloss B, Rosenfeld MG, Heyman RA, Glass CK (1994) Regulation of retinoid signalling by receptor polarity and allosteric control of ligand binding. Nature 371:528–531CrossRefPubMedGoogle Scholar
  20. 20.
    Sanguedolce MV, Leblanc BP, Betz JL, Stunnenberg HG (1997) The promoter context is a decisive factor in establishing selective responsiveness to nuclear class II receptors. EMBO J 16:2861–2873CrossRefPubMedGoogle Scholar
  21. 21.
    IJpenberg A, Tan NS, Gelman L, Kersten S, Seydoux J, Xu J, Metzger D, Canaple L, Chambon P, Wahli W, Desvergne B (2004) In vivo activation of PPAR target genes by RXR homodimers. EMBO J 23:2083–2091CrossRefPubMedGoogle Scholar
  22. 22.
    Kliewer SA, Umesono K, Noonan DJ, Heyman RA, Evans RM (1992) Convergence of 9-cis retinoic acid and peroxisome proliferator signalling pathways through heterodimer formation of their receptors. Nature 358:771–774CrossRefPubMedGoogle Scholar
  23. 23.
    Muerhoff AS, Griffin KJ, Johnson EF (1992) The peroxisome proliferator-activated receptor mediates the induction of CYP4A6, a cytochrome P450 fatty acid omega-hydroxylase, by clofibric acid. J Biol Chem 267:19051–19053PubMedGoogle Scholar
  24. 24.
    DiRenzo J, Soderstrom M, Kurokawa R, Ogliastro MH, Ricote M, Ingrey S, Horlein A, Rosenfeld MG, Glass CK (1997) Peroxisome proliferator-activated receptors and retinoic acid receptors differentially control the interactions of retinoid X receptor heterodimers with ligands, coactivators, and corepressors. Mol Cell Biol 17:2166–2176PubMedGoogle Scholar
  25. 25.
    Shaw N, Elholm M, Noy N (2003) Retinoic acid is a high affinity selective ligand for the peroxisome proliferator-activated receptor beta/delta. J Biol Chem 278:41589–41592CrossRefPubMedGoogle Scholar
  26. 26.
    Jiang YJ, Xu TR, Lu B, Mymin D, Kroeger EA, Dembinski T, Yang X, Hatch GM, Choy PC (2003) Cyclooxygenase expression is elevated in retinoic acid-differentiated U937 cells. Biochim Biophys Acta 1633:51–60PubMedGoogle Scholar
  27. 27.
    Hunter J, Kassam A, Winrow CJ, Rachubinski RA, Capone JP (1996) Crosstalk between the thyroid hormone and peroxisome proliferator-activated receptors in regulating peroxisome proliferator-responsive genes. Mol Cell Endocrinol 116:213–221CrossRefPubMedGoogle Scholar
  28. 28.
    Keller H, Givel F, Perroud M, Wahli W (1995) Signaling cross-talk between peroxisome proliferator-activated receptor/retinoid X receptor and estrogen receptor through estrogen response elements. Mol Endocrinol 9:794–804CrossRefPubMedGoogle Scholar
  29. 29.
    Strickland S, Mahdavi V (1978) The induction of differentiation in teratocarcinoma stem cells by retinoic acid. Cell 15:393–403CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  1. 1.Department of Biomedical Science and Technology, Institute of Biomedical Science and Technology, IBST, RCTCKonkuk UniversitySeoulKorea
  2. 2.Department of Molecular Pharmacology and Experimental TherapeuticsMayo Clinic College of MedicineRochesterUSA

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