Details

Autor(en) / Beteiligte

• Foray, C. F. Arlett, E. P. Malaise, N.

Titel

Underestimation of the small residual damage when measuring DNA double-strand breaks (DSB): is the repair of radiation-induced DSB complete?

Ist Teil von

• International journal of radiation biology, 1999, Vol.75 (12), p.1589-1595

Ort / Verlag

London: Informa UK Ltd

Erscheinungsjahr

1999

Quelle

Taylor & Francis

Beschreibungen/Notizen

• Purpose: To overcome the underestimation of the small residual damage when measuring DNA double-strand breaks (DSB) as fraction of activity released (FAR) by pulsed-field gel electrophoresis. Materials and methods: The techniques used to assess DNA damage (e.g. pulsed-field gel electrophoresis, neutral elution, comet assay) do not directly measure the number of DSB. The Blocher model can be used to express data as DSB after irradiation at 4° C by calculating the distribution of all radiation-induced DNA fragments as a function of their size. We have used this model to measure the residual DSB (irradiation at 4° C followed by incubation at 37° C) in untransformed human fibroblasts. Results: The DSB induction rate after irradiation at 4° C was 39.1 +/- 2.0Gy -1. The DSB repair rate obtained after doses of 10 to 80Gy followed by repair times of 0 to 24h was expressed as unrepaired DSB calculated from the Blocher formula. All the damage appeared to be repaired at 24h when the data were expressed as FAR, whereas 15% of DSB remained unrepaired. The DSB repair rate and the chromosome break repair rate assessed by premature condensation chromosome (PCC) techniques were similar. Conclusion: The expression of repair data in terms of FAR dramatically underestimates the amount of unrepaired DNA damage. The Blocher model that takes into account the size distribution of radiation-induced DNA fragments should therefore be used to avoid this bias. Applied to a normal human fibroblast cell line, this model shows that DSB repair is never complete.