Plasmid DNA breakage by decay of DNA-associated auger emitters: experiments with 123I/125I-iodoHoechst 33258.

PURPOSE

The Auger emitting isotope 123I has a much shorter half-life (13.2 hours), than 125I, the prototype Auger emitter. Monte Carlo simulations and cell culture studies indicate that decay of 123I covalently incorporated into DNA is about half as effective as 125I in terms of DNA breakage and cytotoxicity. The aim of the present study is to assess the DNA breakage efficacy of 123I that is non-covalently associated with DNA, using the minor groove binding ligand iodoHoechst 33258.

MATERIALS AND METHODS

Plasmid (pBR322) DNA was incubated with mixtures of [123I]- and [125I]-iodoHoechst 33258, and DNA double strand breakage (DSB) assessed by assaying the relative amounts of intact, relaxed and linear plasmid forms separated by agarose gel electrophoresis. This "double-label" approach provides a measure of the ratio of probabilities of DSB formation per decay for these two isotopes, with much higher precision than comparing the absolute probabilities for the individual isotopes, principally because it avoids the requirement to accurately determine the fraction of bound ligand.

RESULTS

Our results indicate that the ratio of DSB probability per decay of 123I to that of 125I is 0.63 +/- 0.03. The ratio does not change much with addition of dimethyl sulfoxide (DMSO) to the incubation mixture--0.65 +/- 0.03. This ratio agrees well with the relative efficiency of the two isotopes reported in theoretical and experimental studies, using various endpoints.

CONCLUSIONS

In considering the possible exploitation of the Auger effect in cancer therapy, the modest decrease in DNA breakage efficacy for 123I compared to 125I might be more than compensated for by the advantage of the much shorter half-life. The 60-day half-life of 125I imposes severe limitations in terms of radiation protection.