Archives of Toxicology

, Volume 85, Issue 5, pp 499–504 | Cite as

Acrolein, an I-κBα-independent downregulator of NF-κB activity, causes the decrease in nitric oxide production in human malignant keratinocytes

  • Ki-Young MoonEmail author
Molecular Toxicology


Acrolein, a reactive electrophilic α, β-unsaturated aldehyde, is known to be an alkylating chemical carcinogen. The effect of acrolein on the activation of NF-κB in human malignant epidermal keratinocytes was examined to elucidate the molecular mechanism associated with this NF-κB-acrolein regulation and its consecutive sequence, nitric oxide (NO) production. Acrolein significantly downregulated the cellular NF-κB activity up to 60% compared with control as well as the lipopolysaccharide (LPS)-induced NO production in a dose response manner at concentrations of 10~30 μM. To investigate the regulatory mechanism associated with this NF-κB-acrolein downregulation, the relative level of phosphorylation of I-κBα (serines-32 and -36), a principle regulator of NF-κB activation, represented by acrolein, was quantified. Acrolein inhibited NF-κB activity without altering cellular levels of the phosphorylated and nonphosphorylated forms of I-κBα, implying that the downregulatory effect of acrolein on cellular NF-κB activity in human skin cells is an I-κBα-independent activation pathway. The results suggests that acrolein causes the decrease in nitric oxide production as an I-κBα-independent downregulator of NF-κB activity in human malignant keratinocytes, and acrolein-induced carcinogenesis may be associated with the modulation of cellular NF-κB activity.


Acrolein NF-κB activity I-κB Nitric oxide Chemical carcinogenesis Human malignant keratinocytes 


  1. Baeuerle PA, Baltimore D (1988) I kappa B: a specific inhibitor of the NF-kappa B transcription factor. Science 242:540–546PubMedCrossRefGoogle Scholar
  2. Bécherel PA, Le Goff L, Ktorza S, Ouaaz F, Mencia-Huerta JM, Dugas B, Debré P, Mossalayi MD, Arock M (1995) Interleukin-10 inhibits IgE-mediated nitric oxide synthase induction and cytokine synthesis in normal human keratinocytes. Eur J Immunol 25:2992–2995PubMedCrossRefGoogle Scholar
  3. Bécherel PA, Chosidow O, Le Goff L, Francès C, Debré P, Mossalayi MD, Arock M (1997) Inducible nitric oxide synthase and proinflammatory cytokine expression by human keratinocytes during acute urticaria. Mol Med 3:686–694PubMedGoogle Scholar
  4. Biswal S, Acquaah-Mensah G, Datta K, Wu X, Kehrer JP (2002) Inhibition of cell proliferation and AP-1 activity by acrlein in human A549 lung adenocarcinoma cells due to thiol imbalance and covalent modifications. Chem Res Toxicol 15:180–186PubMedCrossRefGoogle Scholar
  5. Bomsztyk K, Rooney JW, Iwasaki T, Rachie NA, Dower SK, Sibley CH (1991) Evidence that interleukin-1 and phorbol esters activate NF-κB by different pathways: role of protein kinase C. Cell Regul 2:329–335PubMedGoogle Scholar
  6. Bours V, Dejardin E, Goujon-Letawe F, Merville MP, Castronovo V (1994) The NF-kappa B transcription factor and cancer: high expression of NF-kappa B-and I kappa B-relared proteins in tumor cell lines. Biochem Pharmacol 47:145–149PubMedCrossRefGoogle Scholar
  7. Bours V, Bentires-Alj M, Hellin A-C, Viatour P, Robe P, Delhalle S, Benoit V, Merville M-P (2000) Nuclear factor-κB, cancer, and apoptosis. Biochem Pharmacol 60:1085–1090PubMedCrossRefGoogle Scholar
  8. Heck DE, Laskin DL, Gardner CR, Laskin JD (1992) Epidermal growth factor suppresses nitric oxide and hydrogen peroxide production by keratinocytes. Potential role for nitric oxide in the regulation of wound healing. J Biol Chem 267:21277–21280PubMedGoogle Scholar
  9. Horton ND, Biswal SS, Corrigan LL, Bratta J, Kehrer JP (1999) Acrolein causes inhibitor κB-independent decreases in nuclear factor κB activation in human lung adenocarcinoma (A549) cells. J Biol Chem 274:9200–9206PubMedCrossRefGoogle Scholar
  10. Huxford T, Huang DB, Malek S, Ghosh G (1998) The crystal structure of the IkappaBalpha/NF-kappaB complex reveals mechanisms of NF-kappa B inactivation. Cell 95:759–770PubMedCrossRefGoogle Scholar
  11. Karin M, Ben-Neriah Y (2000) Phosphorylation meets ubiquitination: the control of NF-κB activity. Annu Rev Immunol 18:621–663PubMedCrossRefGoogle Scholar
  12. Lambert C, Li J, Jonscher K, Yang TC, Reigan P, Quintana M, Harvey J, Freed BM (2007) Acrolein inhibits cytokine gene expression by alkylating cysteine and arginine residues in the NF-kappaB1 DNA binding domain. J Biol Chem 282:19666–19675PubMedCrossRefGoogle Scholar
  13. Lee SH, Lee SY, Son DJ, Lee H, Yoo HS, Song S, Oh KW, Han DC, Kwon BM, Hong JT (2005) Inhibitory effect of 2’-hydroxycinnamaldehyde on nitric oxide production through inhibition of NF-κB activation in RAW 264.7 cells. Biochem Pharmacol 69:791–799PubMedCrossRefGoogle Scholar
  14. Legrand-Poels S, Zecchinon L, Piret B, Schoonbroodt S, Piette J (1997) Involvement of different transduction pathways in NF-κB activation by several inducers. Free Rad Res 27:301–309CrossRefGoogle Scholar
  15. Li L, Hamilton RF Jr, Holian A (1999) Effect of acrolein on human alveolar macrophage NF-kappaB activity. Am J Physiol 277(3 Pt 1):L550–L557PubMedGoogle Scholar
  16. Moncada S, Palmer RMJ, Higgs EA (1991) Nitric oxide: physiology, pathophysiology, and pharmacology. Pharmacol Rev 43:109–140PubMedGoogle Scholar
  17. Moon K-Y (2007) The chemopreventive effect of retinoids on cellular NF-κB activity induced by NMU and NEU in human malignant keratinocytes. Cancer Res Treat 39:82–87PubMedCrossRefGoogle Scholar
  18. Moon K-Y (2010) N-nitroso-N-methylurea and N-nitroso-N-ethylurea induce upregulation of cellular NF-κB activity through protein kinase C-dependent pathway in human malignant keratinocytes. Arch Pharm Res 33:133–139PubMedCrossRefGoogle Scholar
  19. Moon K-Y, Hahn B-S, Lee J, Kim YS (2001) A cell-based assay system for monitoring NF-κB activity in human HaCaT transfectant cells. Anal Biochem 292:17–21PubMedCrossRefGoogle Scholar
  20. Moon K-Y, Lee YJ, Kim YS (2003) Upregulation of cellular NF-κB activity by alkylating carcinogens in human epidermal keratinocytes. Biol Pharm Bull 26:1195–1197PubMedCrossRefGoogle Scholar
  21. Nath RG, Ocando JE, Chung FL (1996) Detection of 1, N2-propanodeoxyguanosine adducts as potential endogenous DNA lesions in rodents and human tissues. Cancer Res 56:452–456PubMedGoogle Scholar
  22. Nathan C, Xie Q-W (1994) Nitric oxide synthases: roles, tolls, and controls. Cell 78:915–918PubMedCrossRefGoogle Scholar
  23. Perez P, Page A, Jorcano JL (2000) Role of phosphorylated p50-NF-κB in the ultraviolet response of mouse skin. Mol Carcinog 27:272–279PubMedCrossRefGoogle Scholar
  24. Rayet B, Gelinas C (1999) Aberrant rel/nfkb genes and activity in human cancer. Oncogene 18:6938–6947PubMedCrossRefGoogle Scholar
  25. Roméro-Graillet C, Aberdam E, Clément M, Ortonne J-P, Ballotti R (1997) Nitric oxide produced by ultraviolet-irradiated keratinocytes stimulates melanogenesis. J Clin Invest 99:635–642PubMedCrossRefGoogle Scholar
  26. Saliou C, Kitazawa M, McLaughlin L, Yang J-P, Lodge JK, Tetsuka T, Iwasaki K, Cillard J, Okamoto T, Packer L (1999) Antioxidants modulate acute solar ultraviolet radiation-induced NF-kappa-B activation in a human keratinocyte cell line. Free Radic Biol Med 26:174–183PubMedCrossRefGoogle Scholar
  27. Scudiero DA, Shoemaker RH, Paull KD, Monks A, Tierney S, Nofziger TH, Currens MJ, Seniff D, Boyd MR (1988) Evaluation of a soluble tetrazolium/formazan assay for cell growth and drug sensitivity in culture using human and other tumor cell lines. Cancer Res 48:4827–4833PubMedGoogle Scholar
  28. Seo SJ, Choi HG, Chung HJ, Hong CK (2002) Time course of expression of mRNA of inducible nitric oxide synthase and generation of nitric oxide by ultraviolet B in keratinocyte cell lines. Br J Dermatol 147:655–662PubMedCrossRefGoogle Scholar
  29. Toledano MB, Leonard WJ (1991) Modulation of transcription factor NF-κB binding activity by oxidation-reduction in vitro. Proc Natl Acad Sci USA 88:4328–4332PubMedCrossRefGoogle Scholar
  30. Traenckner EB, Pahl HL, Henkel T, Schmidt KN, Wilk S, Baeuerle PA (1995) Phosphorylation of human I-kappaB-alpha on serines 32 and 36 controls I kappaB-alpha proteolysis and NF-kappa B activation in response to diverse stimuli. EMBO J 14:2876–2883PubMedGoogle Scholar
  31. Valacchi G, Pagnin E, Phung A, Nardini M, Schock BC, Cross CE, Van Der Vliet A (2005) Inhibition of NF-kappaB activation and IL-8 expression in human bronchial epithelial cells by acrolein. Antioxid Redox Signal 7:25–31PubMedCrossRefGoogle Scholar
  32. Versteeg HH, Nijhuis E, van den Brink GR, Evertzen M, Pynaert GN, van Deventer SJ, Coffer PJ, Peppelenbosch MP (2000) A new phosphospecific cell-based ELISA for p42/p44 mitogen-activated protein kinase (MAPK), p38 MAPK, protein kinase B and cAMP-response-element-binding protein. Biochem J 350:717–722PubMedCrossRefGoogle Scholar

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© Springer-Verlag 2010

Authors and Affiliations

  1. 1.Department of Clinical PathologyGwangju Health College UniversityGwangjuKorea

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