Camptothecin sensitivity in Werner syndrome fibroblasts as assessed by the COMET technique

J.E. Lowe, Angela Sheerin, Katrin Jennert-Burston, D. Burton, Elizabeth Ostler, J. Bird, M.H.L. Green, Richard Faragher

Research output: Contribution to journalArticle

Abstract

Werner syndrome (WS) is an inherited genetic disease in which individuals display the premature aging of a selected subset of tissues. The disorder results from the loss of function mutations in the wrn gene. Wrn codes for a member of the RecQ helicase family with a unique nuclease domain. There is significant evidence that the role of wrn is to assist in the repair and reinitiation of DNA replication forks that have stalled. Loss of the wrn helicase imposes a distinct set of phenotypes at the cellular level. These include premature replicative senescence (in a subset of cell types), chromosomal instability, a distinct mutator phenotype, and hypersensitivity to a limited number of DNA damaging agents. Unfortunately, most of these phenotypes are not suitable for the rapid assessment of loss of function of the wrn gene product. However, WS cells have been reported to show abnormal sensitivity to the drug camptothecin (an inhibitor of topoisomerase type I). A rapid assay for this sensitivity would be a useful marker of loss of wrn function. The COMET (single-cell gel electrophoresis) assay is a rapid, sensitive, versatile, and robust technique for the quantitative assessment of DNA damage in eukaryotic cells. Using this assay, we have found that a significantly increased level of strand breaks can be demonstrated in WS cells treated with camptothecin compared with normal controls.
Original languageEnglish
Pages (from-to)256-259
Number of pages4
JournalAnnals of the New York Academy of Sciences
Volume1019
DOIs
Publication statusPublished - Jun 2004

Keywords

  • replicative senescence
  • Werner syndrome
  • COMET assay
  • camptothecin

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