Analysis of UV-B-induced DNA damage and its repair in heat-shocked skin cells.
The heat-shock response is a cellular defence mechanism against environmental stresses that is evolutionarily conserved from bacteria to man. Numerous reports demonstrate the beneficial effects of heat-shock protein induction on cell survival under toxic or oxidative stress, e.g., in cardiac and cerebral ischemia or prior to organ transplantation. However, there is little data on the effects of heat treatment on damage caused by UV irradiation. Applying three independent techniques, we have tested the influence of thermal pretreatment of skin cells (1 h, 43 degrees C) on the initial extent of UV-B-induced DNA damage and its subsequent repair. For cultured human epidermal keratinocytes and dermal fibroblasts we can show reduced levels of nucleotide-excision-repair-associated DNA strand incision in the comet assay. Moreover, immunostaining and flow cytometric quantitation of thymidine dimers immediately and one day after irradiation, respectively, reveal that the initial DNA damage is not (keratinocytes) or only moderately (fibroblasts) lower in heat-shocked cells as compared to untreated controls. However, excision repair of dimers is significantly attenuated during the first 24 h in both cell types. Furthermore, using a modified host-cell reactivation assay, we are able to demonstrate that repair of UV-B-damaged plasmid DNA is lower if the transfected cells are previously heat shocked. In summary, heat treatment (1 h, 43 degrees C) inducing heat-shock proteins reduces nucleotide excision repair of UV-B-mediated DNA lesions in fibroblasts and keratinocytes during the following 24 h. This is not necessarily caused by elevated heat-shock protein levels themselves. Possibly the direct thermal damage of repair enzymes is more severe than the potential protective effects of heat-shock proteins.
Schmidt-Rose
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Paul Gerson Unna-Skin Research Center, Beiersdorf AG, Hamburg, Germany. schmidt@hamburg.beiersdorf.com
Pollet
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Will
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Bergemann
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Wittern
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D000328:Adult; D000818:Animals; D002478:Cells, Cultured; D004249:DNA Damage; D004260:DNA Repair; D005347:Fibroblasts / Q000528:radiation effects; D005434:Flow Cytometry; D050884:HSP72 Heat-Shock Proteins; D006360:Heat-Shock Proteins / Q000032:analysis; D006358:Hot Temperature; D006801:Humans; D015603:Keratinocytes / Q000528:radiation effects; D008297:Male; D051379:Mice; D012867:Skin / Q000528:radiation effects*; D014466:Ultraviolet Rays
D050884:HSP72 Heat-Shock Proteins; D006360:Heat-Shock Proteins