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J Biol Chem, Vol. 275, Issue 19, 14684-14690, May 12, 2000

Enhanced DNA Binding and Activation of Transcription Factors NF-kappa B and AP-1 by Acetaldehyde in HEPG2 Cells*

Juan Román, América Giménez, José Maria Lluis, Marta Gassó, Mireia Rubio, Joan Caballeria, Albert Parés, Joan Rodés, and José C. Fernández-ChecaDagger

From the Liver Unit, Institut Malalties Digestives and Instituto de Investigaciones Biomedicas August Pi Suñer, Consejo Superior de Investigaciones Científicas, Barcelona 08036, Spain

Because transcription factors NF-kappa B and activator protein-1 (AP-1) are known to regulate gene expression, we have analyzed the role of acetaldehyde in the activation of NF-kappa B and AP-1 in HepG2 cells. Binding activity and transactivation of NF-kappa B and AP-1 were determined by gel retardation assays and transfection of a luciferase reporter construct controlled by kappa B and AP-1 binding sites, respectively. Acetaldehyde enhanced the DNA binding of NF-kappa B and AP-1 by 1 and 4 h, respectively, increasing the kappa B- and AP-1-dependent luciferase expression. Supershift assays revealed the presence of NF-kappa B heterodimers p65/p50 and p50/p52, whereas nuclear c-Jun levels correlated with the DNA binding of AP-1. The enhanced binding of NF-kappa B to DNA by acetaldehyde in intact cells was accompanied by the proteolytic degradation of Ikappa B-alpha . However, the addition of acetaldehyde to cytostolic extracts from untreated Hep G2 cells did not affect the DNA binding of AP-1 but activated the NF-kappa B heterodimer p65/p50 in the absence of Ikappa B-alpha degradation. Preincubation of HepG2 cells with protein kinase C inhibitors abolished the enhanced DNA binding of NF-kappa B and AP-1 caused by acetaldehyde. Hence, these findings uncover a previously unrecognized role for acetaldehyde in the activation of NF-kappa B and AP-1, which may be of relevance in the alcohol-induced liver disease.

* This work was supported by U.S. National Institute of Alcohol Abuse and Alcoholism Grant AA 09526, Alcohol Center Grant AA11999-01, Dirección General Política Científica y Técnica Grant PM 95-0185, Plan Nacional de I+D Grants SAF 97-0087-C1 and SAF 99-0138, Fondo Investigaciones Sanitarias, Fondo Investigaciones Sanitarias Grant 95-0485, and Europharma.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Dagger To whom correspondence should be addressed: Liver Unit Hospital Clínic i Provincial, Villarroel 170, Barcelona 08036, Spain. Tel.: 34-3-2275709; Fax: 34-3-4515272; E-mail: checa@medicina.ub.es.

Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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