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M. N. Murg M. Schuler D. A. Eastmond 《Mutation Research - Genetic Toxicology and Environmental Mutagenesis》1999,446(2):12
Multicolor fluorescence in situ hybridization (FISH) with tandem-labeling probes for the 1cen-q12 region is a potential biomarker for the detection of structural chromosomal aberrations (CAs) in human cells. To determine the suitability of this technique for biomonitoring humans exposed to 1,3-butadiene (BD) and to characterize the alterations induced as well as their stability over time, the human lymphoblastoid cell line AZH-1 was treated with 5 μM diepoxybutane (DEB) or the positive control mitomycin C (MMC; 0.1 μM) for 24 h. Following the removal of the test chemicals, cell cultures were grown for an additional 19 days in the absence of the test compound. Using the tandem FISH technique, aliquots from the main cultures were examined for the induction of CAs affecting the 1cen-q12 region at various intervals. A significant increase in chromosomal breakage/exchanges affecting the 1cen-q12 region was seen in both the DEB- and MMC-treated interphase and metaphase cells. The damage peaked at approximately 48 h following the addition of the test compound and declined with time. However, at day 20, the frequency of aberrant cells was still significantly higher than the control levels. For comparison, the frequency of micronuclei (MN) formed and their origin was determined using the cytochalasin B-modified MN assay and FISH with a pancentromeric probe. Showing a similar pattern, the frequency of centronere-negative MN peaked at 48 h, but however was not significantly elevated above control levels at 20 days. At early time points, aberrations detected using the FISH assay consisted of nearly equal proportions of unstable- and stable-type aberrations, while at the later time points, translocations were the predominant aberration type. In addition, the use of tandem-label FISH in combination with BrdU-immunfluorescence staining, showed that almost identical frequencies of structural aberrations could be seen in actively replicating and non-replicating cell populations. These studies indicate that a small but significant proportion of the alterations detected using this FISH technique persists over time and that this technique may be valuable for biomonitoring chromosomal alterations in BD-exposed populations. 相似文献
124.
D S Fluck S M Rappaport D A Eastmond M T Smith 《Archives of biochemistry and biophysics》1984,235(2):351-358
1-Naphthol has recently been shown to be selectively toxic to short-term organ cultures of human colorectal tumor tissue. The mechanism underlying 1-naphthol's selective toxicity is as yet unknown, but may be due to the formation of naphthoquinone metabolites, which are known to be highly toxic to tumor cells. By using high-performance liquid chromatography with reductive electrochemical detection, it has been possible to show that 1-naphthol is converted to naphthoquinone metabolites by rat liver microsomes. At least two metabolic pathways, independent of cytochrome P-450, appear to be involved. Iron-dependent lipid peroxidation appears to be responsible for at least part of the conversion of 1-naphthol to predominantly 1,4-naphthoquinone, and it seems likely that superoxide anion radical generation by NADPH-cytochrome P-450 reductase could also catalyze this conversion. 1-Naphthol therefore seems to be converted to cytotoxic naphthoquinone metabolites by mechanism(s) dependent upon the generation of free radicals in rat liver microsomes. The results also demonstrate the utility of HPLC with reductive electrochemical detection for investigations of quinone metabolite formation and the measurement of quinones of both physiological and environmental interest. 相似文献
125.
Two benzene metabolites, catechol and hydroquinone, produce a synergistic induction of micronuclei and toxicity in cultured human lymphocytes 总被引:5,自引:0,他引:5
A mixture of two benzene metabolites, hydroquinone and catechol, produces a striking synergistic genotoxic response in cultured human lymphocytes. This was demonstrated using an anti-kinetochore antibody modification of the micronucleus assay. Treatment with hydroquinone alone or in combination with phenol produced a 3-fold increase in micronucleated cells over background. Treatment with catechol or phenol alone and in combination produced only minor increases in the number of micronucleated cells. In contrast, simultaneous treatment with equimolar (75 microM) concentrations of hydroquinone and catechol resulted in a greater than 16-fold induction of micronucleated cells. Given an additivity model, 20 additional micronucleated cells would be expected (after correcting for background frequencies), yet 140 were observed. Further analysis revealed that over 90% of the micronucleated cells stained positively for kinetochores, indicating a high probability that these micronuclei contain entire chromosomes. This synergistic response appears to occur only at equimolar levels of hydroquinone and catechol. These results suggest that these metabolites are acting together to disrupt the mitotic spindle and interfere with chromosome segregation. These data provide further support for the hypothesis that multiple metabolites acting in concert are involved in the benzene-induced genotoxicity and leukemia in humans. 相似文献