Abstract
The role of UVA-radiation—the major fraction in sunlight—in human skin carcinogenesis is still elusive. We here report that different UVA exposure regime (4 × 5 J/cm2 per week or 1 × 20 J/cm2 per week) caused tumorigenic conversion (tumors in nude mice) of the HaCaT skin keratinocytes. While tumorigenicity was not associated with general telomere shortening, we found new chromosomal changes characteristic for each recultivated tumor. Since this suggested a nontelomere-dependent relationship between UVA irradiation and chromosomal aberrations, we investigated for alternate mechanisms of UVA-dependent genomic instability. Using the alkaline and neutral comet assay as well as γ-H2AX foci formation on irradiated HaCaT cells (20–60 J/cm2), we show a dose-dependent and long lasting induction of DNA single and double (ds) strand breaks. Extending this to normal human skin keratinocytes, we demonstrate a comparable damage response and, additionally, a significant induction and maintenance of micronuclei (MN) with more acentric fragments (indicative of ds breaks) than entire chromosomes particularly 5 days post irradiation. Thus, physiologically relevant UVA doses cause long-lasting DNA strand breaks, a prerequisite for chromosomal aberration that most likely contribute to tumorigenic conversion of the HaCaT cells. Since normal keratinocytes responded similarly, UVA may likewise contribute to the complex karyotype characteristic for human skin carcinomas.
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Acknowledgements
We thank Cees Guikers for the first experiments with daily exposures, Heinrich Steinbauer for performing the tumor experiments, Heidi Holtgreve for her excellent technical assistance in M-FISH hybridization and Stefan Henning for comparing the different UVA-sources. This work was supported in part from the Sander Stiftung, Deutsche Krebshilfe e.V. and EU (LSHC-CT-2004-502943) (both to PB), QLK4-1999-01084 (to PB and KSK) as well as the German Research Foundation (SCHA 411/10-1,-2) and the European Community Marie Curie Program MEIF-CT-2005-023821 (to AR).
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Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc)
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Wischermann, K., Popp, S., Moshir, S. et al. UVA radiation causes DNA strand breaks, chromosomal aberrations and tumorigenic transformation in HaCaT skin keratinocytes. Oncogene 27, 4269–4280 (2008). https://doi.org/10.1038/onc.2008.70
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DOI: https://doi.org/10.1038/onc.2008.70
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