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Processing of Directly and Indirectly Ultraviolet-Induced DNA Damage in Human Cells

  • T. M. Rünger
  • B. Epe
  • K. Möller
Part of the Recent Results in Cancer Research book series (RECENTCANCER, volume 139)

Abstract

Mutations caused by ultraviolet (UV)-induced DNA damage represent the initial genetic changes in the tumorigenesis of UV-induced skin cancer. Different wavelengths of UV radiation cause different kinds of DNA damage and mutations. UVB (290–320 nm) generates pyrimidine dimers by direct excitation of the DNA molecule. UVA (320–400 nm) can damage the DNA only indirectly through a photosensitized reaction. This indirect action is mediated mainly by singlet oxygen, which generates purine base modifications, and has been implicated in the carcinogenic effects of UVA. In order to study the processing of directly and indirectly UV-induced DNA damage in human cells, we first treated the replicating plasmid pRSVcat with up to 10kJ/m2 UVB or with the photosensitizer methylene blue plus visible light (which generates singlet oxygen) in vitro. Then, the damaged plasmid was transfected into normal or repair deficient xeroderma pigmentosum complementation group A (XP-A) cells. DNA repair was assessed by measuring activity of reactivated chloramphenicol acetyltransferase (CAT) enzyme, encoded by the plasmid’s cat gene, in cell extracts after 3 days. While XP-A cells exhibited a significantly reduced repair of UVB-induced DNA damage, they showed a normal repair of singlet oxygen-induced DNA damage. This indicates a differential DNA repair pathway for directly and indirectly UV-induced DNA damage in human cells. Irradiation of the plasmid with UVA alone did not result in a genotoxic effect. Only in conjunction with a cell extract, which provides all candidate cellular photosensitizers, did we find a reduced CAT activity after transfection. This indicates that the genotoxicity of UVA is mediated by a cellular photosensitizer.

Keywords

Singlet Oxygen Xeroderma Pigmentosum Normal Cell Line Chloramphenicol Acetyl Transferase Pyrimidine Dimer 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Springer-Verlag Berlin · Heidelberg 1995

Authors and Affiliations

  • T. M. Rünger
    • 1
  • B. Epe
    • 2
  • K. Möller
    • 1
  1. 1.Department of DermatologyUniversity of WürzburgWürzburgGermany
  2. 2.Department of ToxicologyUniversity of WürzburgWürzburgGermany

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