To clarify the contribution of apoptosis to cell death in four human solid-tumor cell lines, clonogenic cell survival (indicator of radiosensitivity) and induction of caspase-3 and production of DNA fragmentation (markers for apoptosis) were studied in RKO, LS174T, MCF7, and TE671 cells exposed to DNA-incorporated, Auger-electron-emitting 125 I (5-[ 125 I]iodo-2′-deoxyuridine) or γ -radiation. Clonogenic survival was assessed by the colony-forming assay, caspase-3 induction with a fluorogenic substrate, and DNA fragmentation by ligation-mediated PCR. For 125 I, the log dosesurvival curves had no shoulder (high-LET-like) and decreased exponentially at different rates in various cell lines. Induction of caspase-3 in radiosensitive RKO and LS174T cells was 3-fold greater than in radioresistant TE671 and MCF7 cells. Nucleosomal laddering in 125 I-radiosensitive cell lines was dose-dependent, and no laddering was detected in radioresistant lines. For γ -radiation, the survival curve for LS174T cells was monoexponential, and that for the other lines exhibited a distinct shoulder (low-LET-like). The most radiosensitive cell line, LS174T, showed the highest induction of CASP-3, and the most radioresistant line, TE671, the lowest. Although DNA laddering was not detectable in TE671 cells, it was observed in the other lines, being most prominent in LS174T cells. We conclude that apoptosis initiated by DNA-incorporated 125 I is dose-dependent, correlates with radiation sensitivity, and takes place through the CASP-3-mediated pathway, whereas that after γ -irradiation probably occurs via CASP-3-independent and/or CASP-3-mediated pathways and does not correlate with cell radiosensitivity.