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Repeated exposures to UVB induce differentiation rather than senescence of human keratinocytes lacking p16INK-4A

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Abstract

Skin cancers and extrinsic aging are delayed consequences of cumulative UV radiation insults. Exposure of human keratinocytes to UVB has been previously shown to trigger premature senescence. In order to explore the involvement of the cyclin-dependent kinase inhibitor p16INK-4a in UVB-induced premature senescence, we developed an original model of repeated sublethal exposures of human keratinocytes deficient in p16INK-4a. We did not observe any significant increase of senescence-associated β-galactosidase activity positive cells following UVB exposure in this cell line in contrast to primary keratinocytes, suggesting a role for p16INK-4a in UVB-induced senescence. However, we detected sustained DNA damage, prolonged cell cycle arrest, and induction of markers of epidermal differentiation like involucrin and filaggrin as consequences of the repeated exposures. Keratinocytes exposed to the same dose of UVB in a single exposure died. Furthermore, the abundance of the keratins 6, 16 and 17 was increased in keratinocytes exposed repeatedly to UVB suggesting an alternative differentiation. This model allows the induction of a state of differentiation observed in vivo with differentiation uncoupled from premature senescence.

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Abbreviations

CPDs:

Cyclobutane pyrimidine dimers

K:

Cytokeratin

ROS:

Reactive oxygen species

SA β-gal:

Senescence-associated β-galactosidase activity

UVB:

Ultraviolet B

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Acknowledgments

VV is Recipient of a FRIA fellowship. FDC and OT are respectively Post-Doctoral Researcher and Research Associate of the Belgian F·N.R.S. This work received financial support through FRFC Grant 2.4506.01 (Belgium) to YP and OT. We acknowledge the Région Wallonne/FSE for the First-Europe project ‘CosmUV’, the First-DEI project ‘Cosmet-X’, and the ‘Réseaux II Senegene’ and ‘Nanotoxico’ projects. We also thank the European Commission for the Integrated Projects ‘Proteomage’ (LSHM-CT-2005-518230); Coordination Action Link-Age (LSHM-CT-2005-513866); and ‘Matiss’ Marie Curie Project (MTKI-CT-2006–042768).

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Authors and Affiliations

  1. Research Unit of Cellular Biology, Department of Biology, University of Namur (FUNDP), Rue de Bruxelles, 61, 5000, Namur, Belgium

    Véronique Bertrand-Vallery, Emmanuelle Boilan, Noëlle Ninane, Catherine Demazy, Olivier Toussaint & Florence Debacq-Chainiaux

  2. Cell and Tissue Laboratory, URPHYM, University of Namur (FUNDP), Namur, Belgium

    Yves Poumay

  3. Laboratoire de Biologie et Biochimie Cellulaire du Vieillissement, Université Paris 7, Paris, France

    Bertrand Friguet

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  1. Véronique Bertrand-Vallery
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  2. Emmanuelle Boilan
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  3. Noëlle Ninane
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  4. Catherine Demazy
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  5. Bertrand Friguet
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  6. Olivier Toussaint
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  7. Yves Poumay
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  8. Florence Debacq-Chainiaux
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Correspondence to Olivier Toussaint.

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Bertrand-Vallery, V., Boilan, E., Ninane, N. et al. Repeated exposures to UVB induce differentiation rather than senescence of human keratinocytes lacking p16INK-4A . Biogerontology 11, 167–181 (2010). https://doi.org/10.1007/s10522-009-9238-y

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