Mutagenesis, Vol. 14, No. 1, 43-49,
January 1999
© 1999 UK Environmental Mutagen Society/Oxford University Press
Effect of cytochalasin B on the induction of chromosome missegregation by colchicine at low concentrations in human lymphocytes
4 To whom correspondence should be addressed. Tel: +39 6 72594812; Fax: +39 6 2023500; Email: minissi{at}bio.uniroma2.it 1 Dipartimento di Biologia, Università di Roma `Tor Vergata', Viale della Ricerca Scientifica, 00133 Roma, 2 Centro di Genetica Evoluzionistica del CNR, Roma and 3 Dipartimento di Biologia, Università `Roma Tre', Viale Guglielmo Marconi 446, 00146 Roma, Italy
The aim of the present work was to investigate the possible interference of cytochalasin B (cyt B) with low concentration treatment with colchicine in the induction of chromosome/chromatid loss and micronuclei in human lymphocytes mitotically activated in vitro. Thus, cells from a single female donor were treated with colchicine (10 or 25 nM, from 24 h after PHA addition to fixation at 66 h) either in the presence or absence of cyt B. Single lagging chromosomes/chromatids were scored in bipolar ana-telophases and greater damage (disrupted and c-anaphases) was scored in cells at anaphase. Micronuclei were scored in the first 4000 nuclei observed in both cyt B-treated (in mononucleate and binucleate cells) and untreated cultures. With the same criterion, FISH analysis was performed on 2000 nuclei where chromosome 7 and 11 centromeric DNA probes were used in pairs. Our results showed that: (i) the frequency of laggards and of micronuclei increased with colchicine concentration but in the presence of cyt B there was a lower frequency of both (with a mean reduction of ~49%); (ii) FISH analysis showed a colchicine concentration-dependent increase in nuclei with three spots for chromosome 7; (iii) a colchicine concentration-dependent increase in tetraploid cells was observed. This increase was particularly remarkable (5-fold) in cells grown in the presence of cyt B compared with cyt B-untreated cells. The observed `cyt B effects' can be explained if it is assumed that in cytokinesis-blocked cells there is a shorter distance between the poles. As a consequence: (i) laggards would be engulfed in the nearest daughter nucleus with a consequent lower induction of micronuclei; (ii) segregating sister chromatids in heavily impaired anaphases would not travel a sufficient distance to give rise to two daughter nuclei, leading to an increased frequency of polyploid nuclei.
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