Reduction of misleading (“false”) positive results in mammalian cell genotoxicity assays. II. Importance of accurate toxicity measurement

Fowler, Paul; Smith, Robert; Smith, Katie; Young, Jamie; Jeffrey, Laura; Kirkland, David; Pfuhler, Stefan; Carmichael, Paul L.

doi:10.1016/j.mrgentox.2012.04.013

Cited by 73 publications

(48 citation statements)

References 19 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…However, Fowler et al did observe small concentration-related increases in MN frequency that did not reach statistical significance. In addition, in a follow-up publication [31], statistically-significant increases in MN were observed in TK6 cells treated with resorcinol at concentrations of 600, 700 and 1100 g/mL in the presence of cytochalasin B. As such the data from this present study were considered representative.…”

Section: Resorcinol (108-46-3)mentioning

confidence: 77%

“…RLU was calculated per 1000 cells and an increase in caspase 3/7 activity was presented as a fold change relative to the solvent control [31]. Statistical analysis from the data from the followup experiment in the absence of cytochalsin-B was performed as described below.…”

Section: Discussionmentioning

confidence: 99%

“…Previous work [31] has suggested that the use of replication index can underestimate cytotoxicity compared to population doubling with similar chemicals. As no direct side to side comparison of the different treatment conditions (+/− cytochalasin B) was conducted, it is not appropriate to comment further on these observations which were outside the primary aim of these cell comparison investigations.…”

Section: Summary Of Mn Experiments (Reference Tables 5 and 6)mentioning

confidence: 99%

See 2 more Smart Citations

Relationships between p53 status, apoptosis and induction of micronuclei in different human and mouse cell lines in vitro: Implications for improving existing assays

Whitwell

Smith

Jenner

et al. 2015

Mutation Research/Genetic Toxicology and Environmental Mutagene

Self Cite

View full text Add to dashboard Cite

Section: Resorcinol (108-46-3)mentioning

confidence: 77%

Section: Discussionmentioning

confidence: 99%

Section: Summary Of Mn Experiments (Reference Tables 5 and 6)mentioning

confidence: 99%

See 1 more Smart Citation

Relationships between p53 status, apoptosis and induction of micronuclei in different human and mouse cell lines in vitro: Implications for improving existing assays

Whitwell

Smith

Jenner

et al. 2015

Mutation Research/Genetic Toxicology and Environmental Mutagene

Self Cite

View full text Add to dashboard Cite

“…Generally, it is recommended that cells from tissues/species of interest are used (i.e., liver cells to investigate hepatotoxicity and renal cells to predict nephrotoxicity), assuming the cell line or model best represent the in vivo situation. Some cell lines can infl uence assay outputs as shown by Fowler et al [ 22 ] who suggested that the use of human p-53 competent cell lines may reduce the incidence of false positives in genotoxicity assays. Cell lines with reduced or no metabolic capability (e.g., HepG2 cells) may reduce the ability to detect pro-toxicants that undergo metabolic transformation to produce toxic metabolites [ 23 ].…”

Section: Mechanistic Hcs Assaysmentioning

confidence: 99%

High-Content Screening: Understanding and Managing Mechanistic Data to Better Predict Toxicity

Walker

Smith

Frost

et al. 2015

Methods in Pharmacology and Toxicology

View full text Add to dashboard Cite

An increased understanding of the cellular pathways involved in toxicity responses, coupled with a simultaneous advance in technology, has allowed for a shift in the way that the safety assessment of novel chemicals is performed. The development of assays that offer a high-throughput and low-cost option in comparison to more traditional approaches has been a focus of recent years.Early identifi cation of compounds which have the potential to cause Drug Induced Liver Injury (DILI) remains a major challenge for the pharmaceutical industry. Improvements in the mechanistic understanding of the cellular processes involved in this complex response have allowed for models to be generated and more reliable predictions to be made.High-Content Screening (HCS) describes an approach whereby multiple end points can be monitored in a single assay. The focus of this chapter is to introduce HCS with relation to using automated fl uorescence microscopy in order to assess phenotypic changes within cells and, more specifi cally, how this can be incorporated into the drug discovery process.We discuss the advantages that HCS can offer, whilst highlighting important factors to take into account when considering establishing the approach within a laboratory. Four papers whereby HCS has been used to highlight the potential of compounds to cause DILI are reviewed and compared. In addition, an option for including HCS as part of a wider workfl ow to identify environmental toxicants is introduced.

show abstract

“…On the other hand, a miniaturized flow-cytometry based MN test was recently developed to detect chromosome damage, and it is now well proven that this method is a high-throughput technique capable of demonstrating both aneugenic and clastogenic effects simultaneously [16]. Given that p53 deficiency and inaccurate toxicity measurement are the two major reasons resulting in the seemingly high rate of false or misleading positive results, p53-proficient human hepatic cell lines (Hep G2) and more sensitive measurements of toxicity are better choices [17,18]. For this reason information on the two levels of DNA damage and chromosomal damage, including clastogenicity and aneugenicity, can be obtained to provide a comprehensive assessment of the genotoxic potential of source waters.…”

Section: Introductionmentioning

confidence: 99%

Assessing of genotoxicity of 16 centralized source-waters in China by means of the SOS/umu assay and the micronucleus test: Initial identification of the potential genotoxicants by use of a GC/MS method and the QSAR Toolbox 3.0

Yan

Jiang²,

Na³

et al. 2014

Mutation Research/Genetic Toxicology and Environmental Mutagene

View full text Add to dashboard Cite

Only few studies were conducted to assess genotoxicity of centralized source waters in China and almost none of them dealt with the causal relationship between the genotoxic effect and genotoxicants. In this work, 16 centralized source waters in China were sampled from five river systems and genotoxicity of their organic extracts was assessed by use of the SOS/umu test for DNA-damaging effect and the miniaturized flow cytometry-based micronucleus (MN) test for chromosome-damaging effect. In addition, initial identification of potential genotoxicants for the six samples from the Yangtze River was done with a GC/MS method and the QSAR toolbox 3.0. The results demonstrate that eight samples showed both indirect and direct DNA-damaging effects, another four samples showed only indirect DNA-damaging effects, while chromosome-damaging effects were found for 14 out of the 16 samples, in which aneugenic and clastogenic modes of action were found for 4 and 10 samples, respectively. Both direct/indirect DNAdamaging effects and chromosome-damaging effects were induced by the six Yangtze River samples, and the existing different types of genotoxicant confirmed the results. Furthermore, o-phenylphenol was initially identified as the major cause for the DNA-damaging effects while PAHs, pesticides, phenol and anthraquinone were identified as ubiquitous chromosome-damaging agents among these samples. In conclusion, a combination of the SOS/umu test and the miniaturized flow cytometry-based MN test to detect both DNA-damaging and chromosome-damaging effects could be used as a comprehensive genotoxicity assessment tool for the evaluation and classification of genotoxicity of complex mixtures, and potential genotoxicants can be initially identified with additional information from chemical analysis and the QSAR toolbox.

show abstract

Reduction of misleading (“false”) positive results in mammalian cell genotoxicity assays. II. Importance of accurate toxicity measurement

Cited by 73 publications

References 19 publications

Relationships between p53 status, apoptosis and induction of micronuclei in different human and mouse cell lines in vitro: Implications for improving existing assays

Relationships between p53 status, apoptosis and induction of micronuclei in different human and mouse cell lines in vitro: Implications for improving existing assays

High-Content Screening: Understanding and Managing Mechanistic Data to Better Predict Toxicity

Assessing of genotoxicity of 16 centralized source-waters in China by means of the SOS/umu assay and the micronucleus test: Initial identification of the potential genotoxicants by use of a GC/MS method and the QSAR Toolbox 3.0

Contact Info

Product

Resources

About