125I-induced DNA Double Strand Breaks: Use in Calibration of the Neutral Filter Elution Technique and Comparison with X-ray Induced Breaks

Summary

The neutral filter elution assay, for measurement of DNA double strand breakage, has been calibrated using mouse L cells and Chinese hamster V79 cells labelled with [¹²⁵I]dUrd and then held at liquid nitrogen temperature to accumulate decays. The basis of the calibration is the observation that each ¹²⁵I decay, occurring the DNA, produces a DNA double strand break. Linear relationships between ¹²⁵I decays per cell and lethal lesions per cell (minus natural logarithm survival) and the level of elution, were found. Using the calibration data, it was calculated that the yield of DNA double strand breaks after X-irradiation of both cell types was from 6 to 9 × 10^?12 DNA double strand breaks per Gy per dalton of DNA, for doses greater than 6 Gy. Neutral filter elution and survival data for X-irradiated and ¹²⁵I-labelled cells suggested that the relationships between lethal lesions and DNA double strand breakage were significantly different for both cell types. An attempt was made to study the repair kinetics for ¹²⁵I-induced DNA double strand breaks, but was frustrated by the rapid DNA degradation which occurs in cells that have been killed by the freezing-thawing process.