Metabolic activation by a supernatant from human hepatoma cells: a possible alternative in mutagenic tests

Bogaert, M. Duverger-van; Dierickx, Paul J.; Stecca, Claudia; Crutzen, Marc

doi:10.1016/0165-1161(93)90148-s

Cited by 16 publications

(3 citation statements)

References 26 publications

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“…For this reason, indicator cells competent in xenobiotic metabolism would be desirable (Glatt et al, 1990). HepG2 cells are a highly differentiated human hepatoma cell line, which has retained many of the specialized functions usually lost upon culturing and as such represent a suitable in vitro system for studying drug metabolism and (geno)toxicity in man (Duverger-van Bogaert et al, 1993). The other chosen indicator cell line was Caco-2.…”

Section: Resultsmentioning

confidence: 99%

Toxic and genotoxic potential evaluation of soil samples by bioassays

LAH,

AVBERŠEK,

GORJANC

et al. 2005

aas

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Soil samples were obtained from Slovenian industrial region Šaleška dolina and aqueous leachates were prepared. The toxicity and genotoxicity potential of these aqueous fractions have been evaluated by in vitro toxicity and genotoxicity bioassays. Freshwater toxicity test (PROTOXKIT FTM) with a ciliate protozoan Tetrahymena thermophila, and genotoxicity comet assay using Tetrahymena thermophila, Caco-2 and HepG2 cell lines were performed. Biological data were completed through chemical analyses. It has been shown that physico-chemical analyses alone may not be sufficient to characterize soil hazards. To study soil ecotoxcity it is therefore necessary to take into consideration both, the physico-chemical analyses, toxicity and genotoxicity assays.

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Section: Resultsmentioning

confidence: 99%

Toxic and genotoxic potential evaluation of soil samples by bioassays

LAH,

AVBERŠEK,

GORJANC

et al. 2005

aas

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“…Noninduced cells were treated in the same way. The cell pellet was suspended in 10 volumes sucrose-imidazole buffer (0.25 M, pH 7.4) (Duverger- van Bogaert et al, 1993). Cells were homogenized (IKA T8 homogenizor) on ice, and after centrifugation at 9000 g for 10 min at 4 • C, the supernatant fraction was collected and stored at -80 • C.…”

Section: Preparation Of Supernatant Fraction Of Fao Cellsmentioning

confidence: 99%

Evaluation of CYP1A Protein Induction, Biotransformation Capacity and DNA Adduct Formation in a Rat Hepatoma Cell Line (Fao), as Biomarkers of Organic Contamination in Environmental Soil Samples

Østby

Sundby

Krøkje

2006

Water Air Soil Pollut

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Induction of cytochrome P4501A (CYP1A) immunopositive protein was evaluated in the rat hepatoma cell line Fao, as a biomarker of organic pollution in extract of environmental soil samples, exposed to different sources and degrees of chemical contamination. Soil samples were collected in one area in Russia (Monchegorsk) and two areas in Southern Norway (Fiskaa and Birkenes). In addition, one reference soil sample was collected in Central Norway (Høylandet). Contents of selected polycyclic aromatic hydrocarbons (PAHs) in the samples were also evaluated. To further evaluate the inducibility of the most potent soil extract (Fiskaa), S9 fraction of Fao cells, pretreated with this extract, was used as an activation system in the Ames Salmonella assay. The DNA adduct forming capacity of the soil extracts, analyzed by the 32 P-postlabeling technique, was also investigated in Fao cells. The Fao cell line has previously been found to be a very sensitive biomonitoring system, that responds to environmentally relevant concentrations of planar model contaminants with increased level of CYP1A immunopositive protein and DNA adducts. In the present study the Fao cell line also showed its potential for use in evaluating the CYP1A inducing potency of environmental samples. All soil extracts induced CYP1A protein in the Fao cells, although the level of induction varied between the soil samples. The Fiskaa soil extract was the most potent CYP1A inducer and this extract also contained the highest level of PAHs. No significant correlation was observed between the level of the total of 16 PAHs and CYP1A protein level. However, a significant correlation was observed between CYP1A protein level and the level of Benzo[a]pyrene (B[a]P), which is a very potent CYP1A inducer. The S9 fraction of pretreated Fao cells activated B[a]P to mutagens in a concentration-dependent relationship, although the response was weak. No DNA adducts were detected in cells exposed to the soil extracts. This demonstrates the necessity of determining several biomarker parameters simultaneously as one single biomarker may fail to respond to potentially harmful compounds.

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“…Some have proposed that human liver S-9 fraction may be a more appropriate matrix to use in the test, because it would be expected that metabolites relevant to human would be more likely generated [Hakura et al, 2004;Ku et al, 2007]. Reports on the use of human-derived metabolic systems in mutagenicity assays have been described [Beaune et al, 1985;Raineri et al, 1986;Duverger-van Bogaert et al, 1993;Johnson et al, 1996]. However, this would also not be without challenges for use in routine mutagenicity testing, because standardizing the system would be difficult, and obtaining sterile material (to avoid contamination of the test with foreign bacteria) can be challenging.…”

Section: Introductionmentioning

confidence: 99%

Comparison of metabolite profiles generated in Aroclor‐induced rat liver and human liver subcellular fractions: Considerations for in vitro genotoxicity hazard assessment

Obach

Dobo

2008

Environ and Mol Mutagen

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Because it is well known that metabolites of chemicals and drugs are frequently the ultimate species responsible for genotoxicity and carcinogenicity, in vitro testing to identify the human genotoxicity hazard potential of new chemicals and drugs routinely utilizes liver S-9 fraction from rats treated with Aroclor 1254 as a system that can generate metabolites. However, it is frequently questioned as to whether such an in vitro metabolite generation system is the most relevant for human risk, or whether the assay would be better served by using a human-derived in vitro system. To address this, 16 common drugs have been examined for profiles of metabolites in Aroclor-induced rat liver S-9 and pooled human liver S-9. Metabolite profiles were compared using high pressure liquid chromatography coupled with ion trap mass spectrometry, in line with ultraviolet or radiometric detection to help make semiquantitative comparisons. Results showed that, with few exceptions, metabolites generated in the human system were also generated in the rat system. Also, in several cases the rat system generated considerably more metabolites, suggesting that there is a potential that positive genotoxicity findings could be caused by metabolites that have no relevance to humans. These findings suggest that when conducting in vitro genotoxicity testing using the Aroclor-induced rat liver S-9 system, knowledge of the metabolite profile in the system is important, and a comparison to the profile generated in human liver S-9 could be of value when interpreting the genotoxicity results.

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Metabolic activation by a supernatant from human hepatoma cells: a possible alternative in mutagenic tests

Cited by 16 publications

References 26 publications

Toxic and genotoxic potential evaluation of soil samples by bioassays

Toxic and genotoxic potential evaluation of soil samples by bioassays

Evaluation of CYP1A Protein Induction, Biotransformation Capacity and DNA Adduct Formation in a Rat Hepatoma Cell Line (Fao), as Biomarkers of Organic Contamination in Environmental Soil Samples

Comparison of metabolite profiles generated in Aroclor‐induced rat liver and human liver subcellular fractions: Considerations for in vitro genotoxicity hazard assessment

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