Kinetochore analysis of micronuclei allows insights into the actions of colcemid and mitomycin C

Rudd, Noreen L.; Williams, Susan E.; Evans, Martin; Hennig, Ursula; Hoar, David I.

doi:10.1016/0165-1218(91)90098-7

Cited by 42 publications

(9 citation statements)

References 25 publications

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“…The predisposition to induced aneuploidy was analysed by using MN assay associated with C-banding and CREST staining in the presence of agents such as the natural antibiotic mitomycin (MMC), the plant flavonoid quercetin (Quer), both in the presence and in the absence of S9-mix, and of cyclophosphamide (CP). This choice was based on the properties of these genotoxicants, especially the radiomimetic characteristics of MMC and Quer that primarily behave as clastogenic but can secondarily induce aneuploidy [11,12]. We have also taken into account the fact that Quer is one of the most common flavonoids in plants, widely distributed in natural foods, consumed by humans in an average of 50 mg per day [13].…”

Section: Introductionmentioning

confidence: 99%

Aneuploidy induced in lymphocytes of parents of trisomic 21 children

Caria

Chaveca

Rueff

2001

Teratog Carcinog Mutagen

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A possible predisposition to aneuploidy in trisomic 21 individuals, their parents, and a control group was evaluated. Peripheral blood lymphocytes from those three groups were used to study the induction of micronuclei (MN) by mitomycin C, cyclophosphamide, and quercetin. Induced MN were further analysed by C-banding and CREST antibody. Trisomic 21 individuals have spontaneous frequencies of MN significantly higher than their parents and the control group. Quercetin without metabolic activation induces MN in trisomic 21 and their parents at a significantly higher level than in control group. The group of the parents of trisomic 21 individuals exhibits higher frequencies of induced MN by mitomycin C and cyclophosphamide than controls. Mitomycin C significantly induced CREST-positive-MN in ten of the sixteen parents evaluated. The results obtained seem to suggest a unique behaviour for the parents of trisomic 21 patients consisting in an increased susceptibility to chromosome loss in the presence of clastogenic genotoxicants, suggesting a higher predisposition to aneuploidy.

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Section: Introductionmentioning

confidence: 99%

Aneuploidy induced in lymphocytes of parents of trisomic 21 children

Caria

Chaveca

Rueff

2001

Teratog Carcinog Mutagen

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“…Spontaneous micronucleated cell frequency in beluga skin fibroblasts (5–11‰) werein the lower rangereported for human skin fibroblasts (3‐22%) taken from healthy, karyotypically normal individuals and cultured for three to 14 passages [23,24]. Mitomycin C is a direct alkylating agent commonly used as a positive control in the MN assay because it is well recognized as a clastogen and may also act as an aneugen [25]. Induction of MN by MMC in beluga fibroblasts was similar to or lower than that reported for Chinese hamster fibroblasts (fourfold increase at 0.1 μg/ml) and human skin fibroblasts (six‐ to eightfold increase at 1 μg/ml and ninefold increase at 2 μg/ml) [25–27].…”

Section: Discussionmentioning

confidence: 99%

“…Mitomycin C is a direct alkylating agent commonly used as a positive control in the MN assay because it is well recognized as a clastogen and may also act as an aneugen [25]. Induction of MN by MMC in beluga fibroblasts was similar to or lower than that reported for Chinese hamster fibroblasts (fourfold increase at 0.1 μg/ml) and human skin fibroblasts (six‐ to eightfold increase at 1 μg/ml and ninefold increase at 2 μg/ml) [25–27].…”

Section: Discussionmentioning

confidence: 99%

Mercury‐induced micronuclei in skin fibroblasts of beluga whales

Gauthier

Dubeau

Rassart

1998

Enviro Toxic and Chemistry

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Abstract-Beluga whales (Delphinapterus leucas) inhabiting the St. Lawrence estuary are highly contaminated with environmental pollutants and have a high incidence of cancer. Environmental contaminants may be partly responsible for the high incidence of cancer observed in this population. DNA damage plays an important role in the development of cancer. The micronuclei assay was used to test the genotoxic potential of mercury compounds in skin fibroblasts of an Arctic beluga whale. Both mercuric chloride (Hg) and methylmercury (MeHg) induced a highly significant (p Ͻ 0.001) dose-response increase of micronucleated cells. Statistically significant increases in micronucleated cells were observed for 0.5, 5, and 20 g/ml Hg and 0.05, 0.5, and 2 g/ml MeHg when compared to control cultures. Concentrations of 0.5, 5, and 20 g/ml Hg induced a two-, three-and fourfold increase of micronucleated cells, respectively. Treatment with MeHg was one order of magnitude more potent in inducing micronuclei and in inhibiting cell proliferation than Hg. Although results of this in vitro study do not imply that mercury compounds are involved in the etiology of cancer in St. Lawrence beluga whales, significant increases in micronuclei frequency were found at low concentrations of MeHg (0.05 and 0.5 g/ml) that are believed to be comparable to concentrations present in certain whales of this population.

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“…The different DSB-inducing mechanisms generate chromosomal rearrangement that leads to chromosome breaks. Thus, the clastogen-induced MN predominantly represent the kphenotype (lacking kinetochores) containing chromosomal fragments (Eastmond and Tucker, 1989;Rudd et al, 1991;Olaharski et al, 2006).…”

Section: Introductionmentioning

confidence: 99%

Differentiation of Aneugens and Clastogens in the In Vitro Micronucleus Test by Kinetochore Scoring Using Automated Image Analysis

Wilde

Queisser

Holz³

et al. 2018

Environ and Mol Mutagen

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The in vitro micronucleus test according to OECD Test Guideline 487 (TG 487) is widely used to investigate the genotoxic potential of drugs. Besides the identification of in vitro genotoxicants, the assay can be complemented with kinetochore staining for the differentiation between clastogens and aneugens. This differentiation constitutes a major contribution to risk assessment as especially aneugens show a threshold response. Thus, a novel method for automated MN plus kinetochore (k+) scoring by image analysis was developed based on the OECD TG 487. Compound‐induced increases in MN frequency can be detected using the cytokinesis‐block (cytochalasin B) method in V79 cells after 24 h in a 96‐well format. Nuclei, MN, and kinetochores were labeled with nuclear counterstain and anti‐kinetochore antibodies, respectively, to score MN in binuclear or multinuclear cells and to differentiate compound‐induced MN by the presence of kinetochores. First, a reference data set was created by manual scoring using two clastogens and aneugens. After developing the automated scoring process, a set of 14 reference genotoxicants were studied. The automated image analysis yielded the expected results: 5/5 clastogens and 6/6 aneugens (sensitivity: 100%) as well as 3/3 non‐genotoxicants (specificity: 100%) were correctly identified. Further, a threshold was determined for identifying aneugens. Based on the data for our internally characterized reference compounds, unknown compounds that induce ≥53.8% k+ MN are classified as aneugens. The current data demonstrate excellent specificity and sensitivity and the methodology is superior to manual microscopic analysis in terms of speed and throughput as well as the absence of human bias. Environ. Mol. Mutagen. 60:227–242, 2019. © 2018 Wiley Periodicals, Inc.

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Kinetochore analysis of micronuclei allows insights into the actions of colcemid and mitomycin C

Cited by 42 publications

References 25 publications

Aneuploidy induced in lymphocytes of parents of trisomic 21 children

Aneuploidy induced in lymphocytes of parents of trisomic 21 children

Mercury‐induced micronuclei in skin fibroblasts of beluga whales

Differentiation of Aneugens and Clastogens in the In Vitro Micronucleus Test by Kinetochore Scoring Using Automated Image Analysis

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