The neutral filter elution technique has been used to examine the relationship between X-ray-induced DNA double-strand breakage (dsb) and lethal lesions. The ratios of the different lesions produced by X-irradiation were varied by irradiation in the presence of different radiomodifiers, and in each case the same linear relationship between lethal lesions and induced DNA dsb was found. This relationship also held for cells given a hyperthermic treatment before irradiation. It is concluded that DNA dsb is probably the lethal lesion induced by ionizing radiation.

X-ray-induced DNA double-strand breakage (dsb) and lethal lesion induction have been examined using normal and transformed fibroblasts of rodent and human origin. DNA dsb induction varied markedly among the different cell types and was found to reflect radiosensitivity. Linear relationships were found between DNA dsb and lethal lesion induction which were not significantly different for each cell type, suggesting a comparable probability of conversion of DNA dsb into a lethal lesion. The implications of these findings for models of cell survival are discussed.

The mode of death induced by gamma-irradiation in a panel of 10 mouse lymphoid or myeloid cell lines was examined. Four of these lines were known to lose viability (membrane integrity) rapidly after irradiation, whilst the others were known to lose viability considerably more slowly. However, based on the criteria of morphology and DNA degradation pattern, all 10 lines showed apoptotic death. The occurrence of apoptosis after irradiation in rapid-dying lymphoid cell lines was consistent with published results, whilst the demonstration of apoptosis in slow-dying lines was unexpected. Cells of the slow-dying lymphoid lines underwent one or more mitoses prior to death, a feature also reported for fibroblastoid cell lines. However, the occurrence of radiation-induced necrosis in fibroblasts suggests that the pathways leading to 'mitotic death' differ between fibroblastoid and lymphoid cell lines.

Computerized video time-lapse (CVTL) microscopy of X-irradiated cultures of cells of the murine lymphoma cell lines ST4 and L5178Y-S and the human lymphoid cell line MOLT-4 demonstrated that these cells exhibit a wide disparity in the timing of induction and execution of radiation-induced cell death that included rapid-interphase apoptosis, delayed apoptosis, and postmitotic apoptosis. ST4 cells that received 2.5 or 4 Gy of X radiation underwent rapid-interphase apoptosis within 2 h. Apoptosis commenced with a 10-20-min burst of membrane blebbing followed by swelling for 2-4 h and cell collapse. No apoptotic bodies were formed. After a dose of 1 Gy, approximately 90% of ST4 cells died by rapid-interphase apoptosis, while the remainder completed several rounds of cell division prior to cell death. Postmitotic death of ST4 cells occurred with the same morphological sequence of events as during rapid-interphase apoptosis induced by doses of 1-4 Gy. In contrast, L5178Y-S and MOLT-4 cells that received 4 Gy underwent apoptosis more slowly, with a complex series of events occurring over 30-60 h. Only 3% of L5178Y-S cells and 24% of MOLT-4 cells underwent apoptosis without attempting cell division. The cells became abnormally large during a long G(2)-phase delay, and then most of the cells (76-97%) attempted to divide for the first or second time at approximately 18-30 h postirradiation. However, either mitosis failed or division was aberrant; i.e., the large cells divided into three or four fragments which eventually fused together. This process was followed by several rounds of complex and unpredictable membrane blebbing, gross distortions of shape, fragmentation-refusion events, and formation of apoptotic bodies, after which the cells collapsed at 36-60 h postirradiation.

We have examined the timing of gamma-irradiation-induced death in relation to cell cycle progression using a panel of mouse lymphoid or myeloid cell lines. Death was found to occur immediately after irradiation ('rapid interphase' death), or after arrest in G2 phase ('delayed interphase' death). In part II of this series of papers we demonstrated the occurrence of radiation-induced apoptosis in all these cell lines. This suggests that different signals can lead to apoptosis in these cell lines. DNA double-strand breakage appeared to directly stimulate the destruction of cell lines susceptible to rapid interphase death, whilst the signal for delayed interphase and mitotic death, whilst the signal for delayed interphase and mitotic death appeared to be chromosomal aberrations. Several of the cell lines showed different timing of death dependent upon the radiation dose used. These differences in the timing of radiation-induced death are shown to be useful indicators of the relative radiosensitivity of haematopoietic cell lines.