Mitotic non-disjunction as a mechanism for in vitro aneuploidy induction by X-rays in primary human cells

A collaborative study of three laboratories compared the induction of aneuploidy by X-rays in human lymphocytes and fibroblasts. The induction of non-disjunction versus chromosome loss by X-rays was investigated using a variety of aneuploidy detection methods. Chromosome loss was determined by fluorescence in situ hybridization (FISH) with pan-centromeric probes in cytochalasin-B-blocked binucleated cells. Chromosome non-disjunction was estimated by FISH with chromosome-specific centromeric probes in binucleated interphase cells. Chromosomes were counted in parallel in lymphocyte metaphase cells; chromosome counts of the whole karyotype and counts of chromosomes 2 and 8 using chromosome paints. A major observation in spontaneous non-disjunction frequencies concerned the clear difference in frequencies observed between the two painted chromosomes in the same primary cells. When cells were irradiated elevated frequencies were observed for all the different cytogenetic endpoints. Although only a small number of the micronuclei were positive for the centromeric signal and presumably contained whole chromosomes, the absolute number %oC+ increased with dose. Higher rates of non-disjunction were found for irradiated cells; in fibroblasts a statistically significant increase was observed at a dose of 0.5 Gy. The detection of hyperdiploidy by means of chromosome counts and chromosome painting revealed an increase from doses of 1 Gy and higher. Comparison of the different methods detecting different endpoints indicates that non-disjunction may be an important mechanism leading to spontaneous and X-ray-induced aneuploidy. The relative radiosensitivity of aneuploidy induction was compared in two types of primary human cells - lymphocytes and fibroblasts. For chromosome loss both cell types showed similar results, whereas for non-disjunction fibroblasts seemed to be more sensitive. However, these differences may reflect a different sensitivity in the scoring methods used.