Identification of aneuploidy‐inducing agents using cytokinesis‐blocked human lymphocytes and an antikinetochore antibody

Abstract

The identification of agents causing aneuploidy in humans, a condition associated with carcinogenesis and birth defects, is currently limited due to the highly skilled and time-consuming nature of cytogenetic analyses. We report the development of a new simple and rapid assay to identify aneuploidy-inducing agents (aneuploidogens). The assay involves the chemical- or radiation-induced formation of micronuclei in cytokinesis-blocked human lymphocytes and the use of an antikinetochore antibody to determine whether the micronuclei contain centromeres--a condition indicating a high potential for aneuploidy. All agents tested produced dose-related increases in the frequency of micronucleated cells. The micronucleated cells induced by the known aneuploidogens--colchicine, vincristine sulfate, and diethylstilbestrol--contained kinetochore-positive micronuclei 92, 87, and 76% of the time, respectively. In contrast, the micronucleated cells induced by the potent clastogens--ionizing radiation and sodium arsenite--contained kinetochore-positive micronuclei only 3 and 19% of the time, respectively. These results indicate that this relatively simple assay can discriminate between aneuploidogens and clastogens and may allow a more rapid identification of environmental and therapeutic agents with aneuploidy-inducing potential.

Cytokinesis-block micronucleus cytome assay

“…It is therefore an indicator of cytostatic effects and, in the case of lymphocytes, it is also a measure of mitogenic response, which is useful as a biomarker of immune function 22 . NDI is calculated according to the method of Eastmond and Tucker 107 . Score 500 viable cells to determine the frequency of cells with 1, 2, 3 or 4 nuclei, and calculate the NDI using the formula NDI ¼ (M 1 + 2M 2 + 3M 3 + 4M 4 )/N, where M 1 -M 4 represent the number of cells with 1-4 nuclei and N is the total number of viable cells scored (excluding necrotic and apoptotic cells).…”

Evaluación del daño en el ADN y vigilancia biológica de la exposición laboral a solventes orgánicos, 2006

“…Por sus propiedades de volatilidad y liposolubilidad, los solventes tienen un especial tropismo por el sistema nervioso central y periférico, producen cuadros clínicos de neurotoxicidad aguda y crónica, y se consideran mielinotóxicos directos, lo cual se refleja en alteraciones clínicas serias que pueden llegar a ser irreversibles; también, tienen la propiedad de interactuar con los ácidos nucleicos y producir efectos a largo plazo, como mutagénesis o carcinogénesis (4,(12)(13)(14)(15)(16)(17)(18).…”

Instabilidade cromossômica induzida por agroquímicos em trabalhadores rurais na região de Passo Fundo, Rio Grande do Sul, Brasil

“…As análises de micronúcleos em linfócitos de sangue periférico de seres humanos e em linhagens de células cultivadas, mostraram ser efetivas para medir danos citogenéticos de agentes com diferentes mecanismos de genotoxicidade (Eastmond & Tucker, 1989;Fenech & Morley, 1985;Norppa et al, 1993b). Várias pesquisas foram realizadas utilizando cultura de linfó-citos de sangue periférico in vitro, para avaliar o efeito de diferentes praguicidas.…”