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First published online 25 January 2005
doi: 10.1242/jcs.01651

Journal of Cell Science 118, 743-758 (2005)
Published by The Company of Biologists 2005
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Research Article

Repeated exposure of human skin fibroblasts to UVB at subcytotoxic level triggers premature senescence through the TGF-ß1 signaling pathway

Florence Debacq-Chainiaux1, Céline Borlon1, Thierry Pascal1, Véronique Royer1, François Eliaers1, Noëlle Ninane1, Géraldine Carrard2, Bertrand Friguet2, Françoise de Longueville3, Sophie Boffe3, José Remacle1 and Olivier Toussaint1,*

1 Laboratory of Biochemistry and Cellular Biology, Department of Biology, University of Namur (FUNDP), Rue de Bruxelles, 61, 5000 Namur, Belgium
2 Laboratoire de Biologie et Biochimie Cellulaire du Vieillissement – EA 3106 – Université Paris 7, France
3 Eppendorf Array Technologies, Rue du Séminaire, 12, 5000 Namur, Belgium

* Author for correspondence (e-mail: olivier.toussaint{at}fundp.ac.be)

Accepted 17 November 2004

Premature senescence of human diploid fibroblasts (HDFs) can be induced by exposures to a variety of oxidative stress and DNA damaging agents. In this study we developed a robust model of UVB-induced premature senescence of skin HDFs. After a series of 10 subcytotoxic (non-proapoptotic) exposures to UVB at 250 mJ/cm2, the so-called biomarkers of senescence were markedly expressed: growth arrest, senescence-associated ß-galactosidase activity, senescence-associated gene overexpression, deletion in mitochondrial DNA. A set of 44 stress- and senescence-associated genes were found to be differentially expressed in this model, among which clusterin/apolipoprotein J (apo J) and transforming growth factor-ß1 (TGF-ß1). Transfection of apo J cDNA provided protection against premature senescence-inducing doses of UVB and other stressful agents. Neutralizing antibodies against TGF-ß1 or its receptor II (TßRII) sharply attenuated the senescence-associated features, suggesting a role for TGF-ß1 in UVB-induced premature senescence. Both the latent and active forms of TGF-ß1 were increased with time after the last UVB stress. Proteasome inhibition was ruled out as a potential mechanism of UVB-induced stress-induced premature senescence (SIPS). This model represents an alternative in vitro model in photoaging research for screening potential anti-photoaging compounds.

Key words: Fibroblasts, senescence, UVB, clusterin, transforming growth factor beta-1, proteasome, protein oxidation




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