Radiobiological characteristics of solid tumours depending on the p53 status of the tumour cells, with emphasis on the response of intratumour quiescent cells

Abstract

Human head and neck squamous cell carcinoma cells transfected with mutant TP53 (SAS/mTP53) or with a neo vector as a control (SAS/neo) were inoculated subcutaneously (s.c.) into both hind legs of Balb/cA nude mice. Mice bearing tumours received 5-bromo-2′-deoxyuridine (BrdU) continuously to label all proliferating (P) cells in the tumours. The mice then received γ-ray irradiation. Another group of mice received a series of test doses of γ-rays while alive or after tumour clamping to obtain hypoxic fractions (HFs) in the tumours. Right after irradiation, the tumour cells were isolated and incubated with a cytokinesis blocker. The micronucleus (MN) frequency in the cells without BrdU labelling (=quiescent (Q) cells) was determined using immunofluorescence staining for BrdU. Meanwhile, 6 h after irradiation, tumour cell suspensions obtained in the same manner were used for determining the frequency of apoptosis in the Q cells. The MN frequency and apoptosis frequency in total (P+Q) tumour cells were determined from the tumours that were not pretreated with BrdU. In total cell populations, SAS/mTP53 cells were more radioresistant than SAS/neo cells in clonogenic survival. Q tumour cells exhibited a significantly lower apoptosis and MN frequency, probably due to their much larger HF, than total cells. In both total and Q cell fractions, SAS/mTP53 cells were less susceptible to apoptosis and more susceptible to micronucleation than SAS/neo cells. Obviously, TP53 status had the potential to influence the radiosensitivity of not only the total cells, but also the Q cells. However, irrespective of the TP53 status, significant differences in radiosensitivity between total and Q tumour cells were consistently observed. From the viewpoint of tumour control as a whole, including intratumour Q tumour cell control, a treatment modality for enhancing the Q cell response has to be considered.