Induction of lethal mutations in experimental tumours after single and fractionated irradiations in vivo

Abstract

Purpose: To investigate the prolonged reduction in cellular viability (lethal mutations) of surviving cells following irradiation of tumours in vivo and to test the effects of fractionation on the expression of lethal mutations. Materials and methods: A mouse mammary carcinoma (CaNT) was treated with single dose or fractionated X-ray treatments in vivo and survival quantified with an in vitro excision assay soon after irradiation and at various times up to 35 days after in vitro propagation of the surviving cells. Results: A dose-dependent reduction in the plating efficiency was observed in cells isolated from irradiated tumours up to 35 days and many cell generations after irradiation. Considerable heterogeneity in plating efficiency was observed in clonal cell lines isolated from individual colonies grown from irradiated tumours. Delayed expression of lethal damage was observed after fractionated irradiation, although recovery of cellular fitness was greater than after irradiation with single doses (reported previously) suggesting that this form of damage is affected by inter-fraction repair. At equi-toxic doses, delayed expression of lethal damage was similar after three compared with two fractions of radiation per day (reported previously). Conclusions: These effects indicate that conventional excision assays of tumour cell viability under-estimate the total lethal damage caused by irradiation and have implications for modelling of the response of tumours to radiotherapy. The effect of fractionation on expression of this type of damage implies the involvement of repair processes. Therefore the repair proficiency may affect the balance between the immediate and delayed reduction of viability in irradiated cells.