Apoptosis may contribute to false-positive results in the in vitro micronucleus test performed in extreme osmolality, ionic strength and pH conditions

Meintières, Sophie; Marzin, Daniel

doi:10.1016/j.mrgentox.2004.02.003

Cited by 22 publications

(16 citation statements)

References 53 publications

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“…The micronuclei (MNi) formation test is the most widely used method for large-scale detection of broken or lagging chromosomes [(12) and references therein]. However, the origins and fates of MNi have not been completely elucidated (48,49) and intra- and inter-laboratory variability in scoring is still common (48), and complicates the development of a standard protocol for quantitative measurement of chromosome loss rates based on the appearance of MNi. It is also noteworthy that the MNi assay does not measure the fraction of drug-arrested cells that undergo mitosis and form viable aneuploid cells.…”

Section: Discussionmentioning

confidence: 99%

Effects of Anticancer Drugs on Chromosome Instability and New Clinical Implications for Tumor-Suppressing Therapies

Lee

Kouprina

et al. 2016

Cancer Research

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Whole-chromosomal instability (CIN), manifested as unequal chromosome distribution during cell division, is a distinguishing feature of most cancer types. CIN is generally considered to drive tumorigenesis, but a threshold level exists whereby further increases in CIN frequency in fact hinder tumor growth. While this attribute is appealing for therapeutic exploitation, drugs that increase CIN beyond this therapeutic threshold are currently limited. In our previous work, we developed a quantitative assay for measuring CIN based on the use of a non-essential human artificial chromosome (HAC) carrying a constitutively expressed EGFP transgene. Here, we used this assay to rank 62 different anticancer drugs with respect to their effects on chromosome transmission fidelity. Drugs with various mechanisms of action such as antimicrotubule activity, histone deacetylase (HDAC) inhibition, mitotic checkpoint inhibition, and targeting of DNA replication and damage responses were included in the analysis. Ranking of the drugs based on their ability to induce HAC loss revealed that paclitaxel, gemcitabine, dactylolide, LMP400, talazoparib, olaparib, peloruside A, GW843682, VX-680, and cisplatin were the top ten drugs demonstrating HAC loss at a high frequency. Therefore, identification of currently used compounds that greatly increase chromosome mis-segregation rates should expedite the development of new therapeutic strategies to target and leverage the CIN phenotype in cancer cells.

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Section: Discussionmentioning

confidence: 99%

Effects of Anticancer Drugs on Chromosome Instability and New Clinical Implications for Tumor-Suppressing Therapies

Lee

Kouprina

et al. 2016

Cancer Research

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“…Because test systems screening for numerical chromosomal effects rely on labor-intensive microscopic assessment, the micronucleus (MNi) formation test is the most widely used method for large-scale detection of broken or lagging chromosomes [7]. However, because the origins and fates of MNi have not been completely elucidated [29,30], intra- and inter-laboratory variability in scoring is still common [31], and complicates the development of a standard protocol for quantitative measurement of chromosome loss rates based on the appearance of MNi. It is also noteworthy that the MNi assay does not measure the fraction of drug-arrested cells that undergo mitosis and form viable aneuploid cells.…”

Section: Discussionmentioning

confidence: 99%

A new assay for measuring chromosome instability (CIN) and identification of drugs that elevate CIN in cancer cells

Lee

Grimes

et al. 2013

BMC Cancer

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BackgroundAneuploidy is a feature of most cancer cells that is often accompanied by an elevated rate of chromosome mis-segregation termed chromosome instability (CIN). While CIN can act as a driver of cancer genome evolution and tumor progression, recent findings point to the existence of a threshold level beyond which CIN becomes a barrier to tumor growth and therefore can be exploited therapeutically. Drugs known to increase CIN beyond the therapeutic threshold are currently few in number, and the clinical promise of targeting the CIN phenotype warrants new screening efforts. However, none of the existing methods, including the in vitro micronuclei (MNi) assay, developed to quantify CIN, is entirely satisfactory.MethodsWe have developed a new assay for measuring CIN. This quantitative assay for chromosome mis-segregation is based on the use of a non-essential human artificial chromosome (HAC) carrying a constitutively expressed EGFP transgene. Thus, cells that inherit the HAC display green fluorescence, while cells lacking the HAC do not. This allows the measurement of HAC loss rate by routine flow cytometry.ResultsUsing the HAC-based chromosome loss assay, we have analyzed several well-known anti-mitotic, spindle-targeting compounds, all of which have been reported to induce micronuclei formation and chromosome loss. For each drug, the rate of HAC loss was accurately measured by flow cytometry as a proportion of non-fluorescent cells in the cell population which was verified by FISH analysis. Based on our estimates, despite their similar cytotoxicity, the analyzed drugs affect the rates of HAC mis-segregation during mitotic divisions differently. The highest rate of HAC mis-segregation was observed for the microtubule-stabilizing drugs, taxol and peloruside A.ConclusionThus, this new and simple assay allows for a quick and efficient screen of hundreds of drugs to identify those affecting chromosome mis-segregation. It also allows ranking of compounds with the same or similar mechanism of action based on their effect on the rate of chromosome loss. The identification of new compounds that increase chromosome mis-segregation rates should expedite the development of new therapeutic strategies to target the CIN phenotype in cancer cells.

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“…More recent research indicates that extreme culture conditions (hypo-and hyperosmolality and high pH) induce apoptosis and necrosis, leading to DNA fragmentation and producing false-positive responses in clastogenic assays (Meintieres and Marzin, 2004).…”

Section: Statusmentioning

confidence: 99%

Scientific Opinion on Flavouring Group Evaluation 67, Revision 1 (FGE.67Rev.1): Consideration of 40 furan-substituted aliphatic hydrocarbons, alcohols, aldehydes, ketones, carboxylic acids and related esters, sulfides, disulfides and ethers evaluated by J

Flavourings¹

2011

EFSA Journal

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The Panel on Food Contact Materials, Enzymes, Flavourings and Processing Aids of the European Food Safety Authority was requested to consider evaluations of flavouring substances assessed since 2000 by the Joint FAO/WHO Expert Committee on Food Additives (the JECFA), and to decide whether further evaluation is necessary, as laid down in Commission Regulation (EC) No 1565/2000. The present consideration concerns a group of 33 furan‐substituted aliphatic hydrocarbons, alcohols, aldehydes, ketones, carboxylic acids and related esters, sulfides, disulfides and ethers evaluated by the JECFA. In the present version of FGE.67 eight additional substances have been included. The substances were evaluated through a stepwise approach (the Procedure) that integrates information on structure‐activity relationships, intake from current uses, toxicological threshold of concern, and available data on metabolism and toxicity. For twenty‐two substances [FL‐no: 13.029, 13.030, 13.045, 13.052, 13.054, 13.059, 13.061, 13.066, 13.069, 13.070, 13.083, 13.092, 13.101, 13.103, 13.105, 13.106, 13.107, 13.123, 13.138, 13.148, 13.163 and 13.191] a concern for genotoxicity was raised and therefore these were not evaluated using the Procedure. The Panel concluded that 8 substances [FL‐no: 13.006, 13.021, 13.022, 13.023, 13.024, 13.074, 13.116 and 13.190] do not give rise to safety concerns at the levels of dietary intake, estimated on the basis of the MSDI approach. For one substance [FL‐no: 13.058] additional toxicity data are requested. Besides the safety assessment of these substances, the specifications for the materials of commerce have been considered. For three substances [FL‐no: 13.031, 13.045 and 13.047] data on specifications / stereoisomerism are missing.

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Apoptosis may contribute to false-positive results in the in vitro micronucleus test performed in extreme osmolality, ionic strength and pH conditions

Cited by 22 publications

References 53 publications

Effects of Anticancer Drugs on Chromosome Instability and New Clinical Implications for Tumor-Suppressing Therapies

Effects of Anticancer Drugs on Chromosome Instability and New Clinical Implications for Tumor-Suppressing Therapies

A new assay for measuring chromosome instability (CIN) and identification of drugs that elevate CIN in cancer cells

Scientific Opinion on Flavouring Group Evaluation 67, Revision 1 (FGE.67Rev.1): Consideration of 40 furan-substituted aliphatic hydrocarbons, alcohols, aldehydes, ketones, carboxylic acids and related esters, sulfides, disulfides and ethers evaluated by J

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