THE CYTOCHALASIN-B MICRONUCLEUS KINETOCHORE ASSAY INVITRO - STUDIES WITH 10 SUSPECTED ANEUGENS

An in vitro micronucleus assay in low passage Chinese hamster Luc2 cells capable of detecting numerical and structural chromosome changes was developed. Chromosome loss was inferred by indirect visualisation of human CREST antikinetochore antibodies bound to centromeres in chemically-induced micronuclei of cytochalasin-B arrested binucleated cells. The assay was used to evaluate 10 chemicals which had been selected for their known or suspected effects upon various components of the cell-division apparatus. These chemicals were colchicine (COL), vinblastine (VBL), thiabendazole (TBZ), chloral hydrate (CH), thimerosal (TM), diazepam (DZ), pyrimethamine (PYR), hydroquinone (HQ), cadmium chloride (CdCl2) and econazole nitrate (EZ). Mitomycin-C (MMC) was used as a positive control for the induction of micronuclei. 8 of the core chemicals induced micronuclei in Chinese hamster Luc2 cells. 4 of the chemicals (COL, VBL, TBZ, CH) increased levels of micronuclei which were positive for kinetochore antibody labelling and hence chromosome loss. 3 of the chemicals (DZ, PYR, HQ) and the positive control (MMC) increased the levels of Mn which were negative for kinetochore antibody labelling. The results with TM were equivocal and EN was negative. The results of these studies suggest that the cytochalasin-B Mn/k assay is a cost-effective, simple and rapid alternative to classical cytogenetic assays for the detection of chemically induced aneuploidy.