Some observations on the localization of mitomycin C-induced aberrations in human lymphocytes

Actively cycling human lymphocytes were treated with mitomycin C for 1 h (1.4 micrograms/ml) and then grown in medium containing 10 micrograms/ml bromodeoxyuridine. Serial 5-h colcemid accumulation samples were taken up to 35 h and the air-dried methaphase spreads stained for replication banding. A complete cell-cycle subphasing analysis was made, and classified cells scored for all categories of chromatid-type aberrations and their location. In spite of the high dose which produced massive delay and cycle perturbation, there was no evidence for selective lethality of early-S cells, in fact such cells were in excess. Extreme localization of aberrations to late-replicating (mostly centromeric) regions was found at all subphases and in pre-S cells. This rules out 'localization by default' as an explanation for the observed preferential occurrence of 'break points' in these regions. The frequency of incomplete intrachanges, low in late S, rises dramatically in early S to become maximal in pre-S cells.