Characterization of radiation sensitivity of human squamous carcinoma A431 cells

Abstract

Studies have been performed to investigate the radiosensitivity of human squamous carcinoma cells. A431 cells were grown in vitro as exponential and fed-plateau monolayer cultures or as multicellular spheroids. Radiobiological studies of various cultures showed that fed-plateau phase cells were more sensitive (D0 = 1.3 Gy) than exponentially growing cells (D0 = 1.5 Gy). After a single dose of 12 Gy or two doses of 6 Gy irradiation, A431 cultures exhibited a large capacity for potentially lethal damage (PLD) repair (PLD repair factor = 17), but a relatively small sublethal damage (SLD) repair. In order to measure the radiation sensitivity of proliferating (P) and quiescent (Q) cells, enriched populations of P- and Q-cells were isolated from A431 spheroids. Flow cytometric analysis with acridine orange (AO) staining demonstrated that there was a shift of the RNA histograms in fed-plateau and spheroid cultures towards lower values, suggesting the presence of a subpopulation of Q-cells. Centrifugal elutriation was used to isolate the Q-cells from dissociated spheroid cells. Coulter cell volume distributions and flow cytometric analysis showed that Q-cells had a small cell volume (approximately 1380 microns3), low RNA content and a G1-like DNA content. Continuous labelling experiments with tritiated thymidine confirmed the non-proliferating nature of the Q-cells. Irradiation of the Q-cells after isolation from spheroids with between 0 to 10 Gy showed that they were more radiosensitive (decreased D0) than the P-cells isolated from these spheroids. The latter were, however, similar in radiosensitivity to exponential G1 cells.