Mutagenesis, Vol. 14, No. 4, 385-389,
July 1999
© 1999 UK Environmental Mutagen Society/Oxford University Press
A proposal for a simple way to distinguish aneugens from clastogens in the in vitro micronucleus test
Division of Genetics and Mutagenesis, National Institute of Health Sciences, 1-18-1 Kamiyoga, Setagaya-ku, Tokyo 158-8501, Japan
In our previous in vitro micronucleus (MN) study, we showed that aneugens, in addition to inducing micronuclei, induce a higher frequency of polynuclear (PN) and mitotic (M) cells than clastogens. We hypothesized that the frequency of PN and M cells induced can distinguish aneugens from clastogens. To test the hypothesis, we conducted the micronucleus tests with mitomycin C (MMC), N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), vincristine (VINC) and diazepam in a Chinese hamster cell line (CHL) and VINC, benzo[a]pyrene (BP) and 7,12-dimethylbenz[a]anthracene (DMBA) in a subclone of V79 cells (V79-MZ). All chemicals increased the frequency of M cells with statistical significance. All chemicals except diazepam increased the frequency of PN cells with statistical significance. Three of the aneugens (VINC, BP and DMBA) induced 
200 PN cells/1000 cells while the clastogens (MNNG and MMC) induced 100 PN cells at most. All the aneugens but no clastogens significantly increased the frequency of M cells. We propose that micronucleus test-positive chemicals that induce 200 PN cells/1000 cells and significantly increase the frequency of M cells are aneugens and those that induce at most 100 PN cells/1000 cells and do not significantly increase the frequency of M cells in our MN test protocol are clastogens. Diazepam, however, did not induce PN cells, although it increased the frequency of M cells dose dependently. We explain this fact in relation to diazepam's mode of action. Our proposal suggests a quick, easy and practical way to distinguish aneugens from clastogens for screening purposes.
1 To whom correspondence should be addressed. Tel: +81 3 3700 1141; Fax: +81 3 3700 2348; Email: matsuoka{at}nihs.go.jp
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