Six human cervical cancer cell lines [five human papillomavirus (HPV) positive, one HPV negative] for induction and rejoining of DNA strand breaks and for kinetics of formation and loss of serine 139 phosphorylated histone H2AX (γH2AX). X-rays induced the same level of DNA breakage for all cell lines. By 8 hours after 20 Gy, <2% of the initial single-strand breaks remained and no double-strand breaks could be detected. In contrast, 24 hours after irradiation, γH2AX representing up to 30% of the initial signal still present. SW756 cells showed almost four times higher background levels of γH2AX and no residual γH2AX compared with the most radiosensitive HPV-negative C33A cells that showed the lowest background and retained 30% of the maximum level of γH2AX. Radiation sensitivity, measured as clonogenic-surviving fraction after 2 Gy, was correlated with the fraction of γH2AX remaining 24 hours after irradiation. A substantial correlation with γH2AX loss half-time measured over the first 4 hours was seen only when cervical cell lines were included in a larger series of p53-deficient cell lines. Interestingly, p53 wild-type cell lines consistently showed faster γH2AX loss half-times than p53-deficient cell lines. We conclude that cell line-dependent differences in loss of γH2AX after irradiation are related in part to intrinsic radiosensitivity. The possibility that the presence of γH2AX foci may not always signify the presence of a physical break, notably in some tumor cell lines, is also supported by these results.