Contribution of lethal mutations to excision assays for tumour cell survival

Conventional assays of cell survival determine only the proportion of colony-forming cells, assuming that all such cells are equivalent. However, cells surviving irradiation are reported to have lower plating efficiencies than unirradiated controls, suggesting an additional component of cellular damage that is ignored in conventional survival assays, but which could contribute to therapeutic outcome. Therefore we have examined the contribution of this additional form of damage to excision assays for cell survival in experimental tumours following both single dose and fractionated irradiation (10F/5 days) in vivo. Plating efficiencies were considerably lower for the long-term descendents of irradiated compared with non-irradiated cells. Expression of delayed reproductive death was reduced after fractionated radiation doses, only appearing after a substantial number of 3.4 Gy fractions had accumulated. Thus estimates of survival derived from single clonogenicity assays may underestimate the reduction in cell viability from a particular treatment. This could compromise assays for intrinsic radiosensitivity and mathematical modelling of the efficacy of treatment regimens.