Aphidicolin Synchronization of Mouse L Cells Perturbs the Relationship between Cell Killing and DNA Double-strand Breakage after X-irradiation

Abstract

Mouse L cells were synchronized in early S-phase by two 12 h incubations in medium containing aphidicolin (2 micrograms/ml), separated by 8 h in drug-free medium. The relationship between X-ray-induced cell killing and DNA double-strand breakage was then examined for cells that had entered S-phase, G2-phase, mitosis, and G1-phase following release from aphidicolin and was compared to the response of asynchronous cultures. Aphidicolin-synchronized cells showed cycle phase-dependent changes in their dose-responses for both killing and DNA dsb. However, on the basis of the level of DNA dsb per unit length of DNA required to produce a lethal lesion, aphidicolin-synchronized cells were more sensitive to X-rays than were asynchronous cultures. This sensitivity peaked 2 h after release from aphidicolin treatment and then progressively declined towards the asynchronous culture value. It is argued that these results are due to deregulation of the temporal order of DNA replication following aphidicolin treatment, and can be incorporated into the critical DNA target size model (Radford, Hodgson, and Matthews, in preparation) by postulating that the targets for radiation action in mammalian cells are DNA-associated with potentially transcriptionally active proto-oncogenes or constitutive fragile sites.