Cytosine arabinoside, vinblastine, 5-fluorouracil and 2-aminoanthracene testing in the in vitro micronucleus assay with L5178Y mouse lymphoma cells at Sanofi Aventis, with different cytotoxicity measurements, in support of the draft OECD Test Guideline on In Vitro Mammalian Cell Micronucleus Test

Cariou, Olivier; Laroche-Prigent, Nathalie; Ledieu, Sandrine; Guizon, Isabelle; Paillard, F.; Thybaud, Véronique

doi:10.1016/j.mrgentox.2010.04.005

Cited by 9 publications

(6 citation statements)

References 7 publications

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“…Taken together, the secondary toxicity and specificity screens identified three macropinocytosis inhibitors (imipramine, phenoxybenzamine and vinblastine) that exert no cytotoxic effects at IC 100 concentrations after a longer exposure and lack inhibitory activities against phagocytosis and clathrin‐mediated and caveolin‐mediated endocytosis. The microtubule inhibitor vinblastine was removed from further analysis as it inhibits important physiological processes, such as cell cycle, and has serious undesired side effects in vivo (Cariou et al ., ).…”

Section: Resultsmentioning

confidence: 97%

“…These three drugs, therefore, were excluded from further analysis. The microtubule inhibitor vinblastine was also excluded from further study as it is known to inhibit the cell cycle at low concentrations and have serious undesired side effects in vivo (Cariou et al ., ). Interestingly, the inhibitory effect of vinblastine on macropinocytosis supports the role of microtubule‐mediated intracellular transport (i.e.…”

Section: Discussionmentioning

confidence: 97%

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Identification of novel macropinocytosis inhibitors using a rational screen of Food and Drug Administration‐approved drugs

Lin

Singla

Ghoshal

et al. 2018

British J Pharmacology

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Section: Resultsmentioning

confidence: 97%

Section: Discussionmentioning

confidence: 97%

Identification of novel macropinocytosis inhibitors using a rational screen of Food and Drug Administration‐approved drugs

Lin

Singla

Ghoshal

et al. 2018

British J Pharmacology

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“…Test chemicals were purchased from MilliporeSigma (Billerica, MA, USA) and were selected since there is significant published literature that demonstrates their potential to induce MN across a wide range of doses 12,13,[59][60][61] . MMC (CAS no.…”

Section: Test Chemicalsmentioning

confidence: 99%

The in vitro micronucleus assay using imaging flow cytometry and deep learning

et al. 2021

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The in vitro micronucleus (MN) assay is a well-established assay for quantification of DNA damage, and is required by regulatory bodies worldwide to screen chemicals for genetic toxicity. The MN assay is performed in two variations: scoring MN in cytokinesis-blocked binucleated cells or directly in unblocked mononucleated cells. Several methods have been developed to score the MN assay, including manual and automated microscopy, and conventional flow cytometry, each with advantages and limitations. Previously, we applied imaging flow cytometry (IFC) using the ImageStream® to develop a rapid and automated MN assay based on high throughput image capture and feature-based image analysis in the IDEAS® software. However, the analysis strategy required rigorous optimization across chemicals and cell lines. To overcome the complexity and rigidity of feature-based image analysis, in this study we used the Amnis® AI software to develop a deep-learning method based on convolutional neural networks to score IFC data in both the cytokinesis-blocked and unblocked versions of the MN assay. We show that the use of the Amnis AI software to score imagery acquired using the ImageStream® compares well to manual microscopy and outperforms IDEAS® feature-based analysis, facilitating full automation of the MN assay.

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“…Automatic scoring of MN was performed using MNScore software (version 3.8.101; MetaSystems) with a Metafer 4 image analyser (MetaSystems) comprising a Zeiss Axioplan 2 imager (Schunck et al, ; Cariou et al, ) to detect MN and an operator validated and excluded the false MN. Two thousand mononucleated cells for cell lines L5178Y and TK6 and two thousand binucleated cells for blood human lymphocytes per culture (two cultures per concentration, i.e.…”

Section: Methodsmentioning

confidence: 99%

Genotoxic risk of ethyl-paraben could be related to telomere shortening

Finot¹,

Kaddour

Morat

et al. 2016

J. Appl. Toxicol.

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The ability of parabens to promote the appearance of multiple cancer hallmarks in breast epithelium cells provides grounds for regulatory review of the implication of the presence of parabens in human breast tissue. It is well documented that telomere dysfunction plays a significant role in the initiation of genomic instability during carcinogenesis in human breast cancer. In the present study, we evaluated the genotoxic effect of ethyl 4-hydroxybenzoate (ethyl-paraben), with and without metabolic activation (S9), in studies following OECD guidelines. We observed a significant increase in genotoxic damage using the Mouse Lymphoma Assay and in vitro micronucleus (MN) tests in the L5178Y cell line in the presence of S9 only after a short exposure. A high frequency of MN was observed in the TK6 cells after a short exposure (3 h) in the presence of S9 and a long exposure (26 h) without S9. We found significant increases in the MN frequency and induced chromosomal aberrations in the lymphocytes of only one donor after ethyl-paraben exposure in the presence of S9 after a short exposure. Cytogenetic characterization of the paraben-treated cells demonstrated telomere shortening associated with telomere loss and telomere deletions in L5178Y and TK6 cells and lymphocytes of the paraben sensitive-donor. In a control cohort of 68 human lymphocytes, telomere length and telomere aberrations were age-dependent and showed high inter-individual variation. This study is the first to link telomere shortening and the genotoxic effect of ethyl paraben in the presence of S9 and raises the possibility that telomere shortening may be a proxy for underlying inter-individual sensitivity to ethyl-paraben. Copyright © 2016 John Wiley & Sons, Ltd.

show abstract

Cytosine arabinoside, vinblastine, 5-fluorouracil and 2-aminoanthracene testing in the in vitro micronucleus assay with L5178Y mouse lymphoma cells at Sanofi Aventis, with different cytotoxicity measurements, in support of the draft OECD Test Guideline on In Vitro Mammalian Cell Micronucleus Test

Cited by 9 publications

References 7 publications

Identification of novel macropinocytosis inhibitors using a rational screen of Food and Drug Administration‐approved drugs

Identification of novel macropinocytosis inhibitors using a rational screen of Food and Drug Administration‐approved drugs

The in vitro micronucleus assay using imaging flow cytometry and deep learning

Genotoxic risk of ethyl-paraben could be related to telomere shortening

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