Chromosomal effects of newly identified water pollutants PBTA-1 and PBTA-2 and their possible mother compounds (azo dyes) and intermediates (non-ClPBTAs) in two Chinese hamster cell lines.
We performed the in vitro micronucleus (MN) test on 2-[2-(acetylamino)-4-[bis(2-methoxyethyl)amino]-5-methoxyphenyl]-5-amino-7-bromo-4-chloro-2H-benzotriazole (PBTA-1) and 2-[2-(acetylamino)-4-[N-(2-cyanoethyl)-ethylamino]-5-methoxyphenyl]-5-amino-7-bromo-4-chloro-2H-benzotriazole (PBTA-2), which are newly identified water pollutants from the Nishitakase river in Kyoto, Japan, and on their possible mother compounds (AZO DYE) and intermediates (non-ClPBTAs). We tested these compounds in the absence and presence of S9 mix in two Chinese hamster cell lines CHL and V79-MZ and scored MN, polynuclear and karyorrhectic (PN), and mitotic (M) cells. PBTA-2 in the absence of S9 mix induced the strongest responses in both cell lines. It was also a strong inducer of binucleate cells in PN cells in both cell lines, which suggested that it induced polyploidy. PBTA-1 showed clear positive results only in the absence of S9 mix and only in V79-MZ cells, inducing aneuploidy. In CHL cells AZO DYE-1 significantly induced MN cells in the presence of S9 mix, and AZO DYE-2 induced MN and PN cells, including binucleate cells and cells with a multilobed nucleus, in the absence of S9 mix. In V79-MZ cells, AZO DYE-1 and -2 induced primarily M cells in the presence of S9 mix. 9% of the M cells treated with 50 microg/ml AZO DYE-1 showed endoreduplication. AZO DYE-2 at 200 microg/ml condensed the chromatin in 100% of the cells. The non-ClPBTAs were a bit more cytotoxic than the other compounds and induced a slight increase in MN cells in both cell lines. Some of the chemicals tested induced a characteristic karyomorphology that might reflect abnormal cell division. Abnormalities of cell division could be detected in PN and M cells as well as in MN cells. Structure-activity relationships have also been discussed.
Matsuoka
A
A
Division of Genetics and Mutagenesis, National Institute of Health Sciences, 1-18-1 Kamiyoga, Setagaya-ku, Tokyo 158-8501, Japan. matsuoka@nihs.go.jp
Tada
A
A
Terao
Y
Y
Nukaya
H
H
Onfelt
A
A
Wakabayashi
K
K
D000818:Animals; D001391:Azo Compounds / Q000302:isolation & purification / Q000633:toxicity; D002460:Cell Line; D002869:Chromosome Aberrations; D004396:Coloring Agents / Q000302:isolation & purification / Q000633:toxicity; D006224:Cricetinae; D066298:In Vitro Techniques; D008297:Male; D015162:Micronucleus Tests; D008862:Microsomes, Liver / Q000378:metabolism; D009153:Mutagens / Q000302:isolation & purification / Q000633:toxicity*; D051381:Rats; D014230:Triazoles / Q000302:isolation & purification / Q000633:toxicity*; D014874:Water Pollutants, Chemical / Q000302:isolation & purification / Q000633:toxicity*
C115463:2-(2-(acetylamino)-4-(N-(2-cyanoethyl)ethylamino)-5-methoxyphenyl)-5-amino-7-bromo-4-chloro-2H-benzotriazole; C112866:2-(2-(acetylamino)-4-(bis(2-methoxyethyl)amino)-5-methoxyphenyl)-5-amino-7-bromo-4-chloro-2H-benzotriazole; D001391:Azo Compounds; D004396:Coloring Agents; D009153:Mutagens; D014230:Triazoles; D014874:Water Pollutants, Chemical