Cytotoxicity, extent of DNA double-strand breaks, and stimulation of poly(adenosine diphosphoribose)-synthesis were measured in two established human head and neck squamous carcinoma cell lines (183A and 1483) following x-irradiation. The 1483 cell line was 15-fold more resistant to x-ray-mediated cytotoxicity than was the 183A cell line. X-ray-mediated DNA strand cleavage also differed in these two cell lines with the absolute frequency of DNA double-strand breaks in the sensitive cells 183A cells being twice that in the resistant 1483 cell line. No detectable stimulation of poly(adenosine diphosphoribose)-synthesis was measured in the sensitive 183A cells whereas a marked increase in incorporation of [3H]-nicotinamide adenine dinucleotide was readily detected following x-irradiation of the resistant 1483 cells. These findings suggest a possible role of altered poly(adenosine diphosphoribose)-synthesis in the sensitivity of human head and neck squamous carcinoma cells to ionizing radiation.