An in vitro rainbow trout cell bioassay for aryl hydrocarbon receptor‐mediated toxins

Richter, Catherine A.; Tieber, Virginia L.; Denison, Michael S.; Giesy, John P.

doi:10.1002/etc.5620160321

Cited by 59 publications

(35 citation statements)

References 46 publications

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“…MVLN cells are MCF‐7 human breast carcinoma cells stably transfected with a luciferase reporter gene under control of estrogen response elements of the Xenopus vitellogenin A2 gene [23]. H4IIE‐luc cells and RLT 2.0 cells are rat and rainbow trout hepatoma cells, respectively, which were stably transfected with a luciferase reporter gene under control of dioxin‐responsive enhancers (DREs) [20,24]. All cells were cultured in 100‐mm disposable petri plates (Corning, Corning, NY, USA) and were incubated in a humidified 95:5 air:CO 2 atmosphere.…”

Section: Methodsmentioning

confidence: 99%

Alkylphenols, polycyclic aromatic hydrocarbons, and organochlorines in sediment from Lake Shihwa, Korea: Instrumental and bioanalytical characterization

Khim

Villeneuve

Kannan

et al. 1999

Enviro Toxic and Chemistry

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Lake Shihwa is an artificial lake, located on the west coast of Korea, that has experienced environmental deterioration since 1994, when it was formed by construction of a sea dike. This study used instrumental analysis and in vitro bioassays to characterize organic contaminants in sediment collected from 11 stations on Lake Shihwa. Alkylphenol (AP) concentrations in Lake Shihwa sediment ranged from 20.2 to 1,820 ng/g nonylphenol and from 4.69 to 50.5 ng/g octylphenol, on a dry weight basis. Maximum concentrations of polycyclic aromatic hydrocarbons (PAHs), organochlorine pesticides, and polychlorinated biphenyls (PCBs) were 30.8, 2.26, and 12.3 ng/g (dry weight), respectively. Significant estrogenic activity was associated with fractions containing APs. Mass-balance analysis suggested that reported concentrations of APs account for less than 20% of the estrogenic activity observed. No significant dioxinlike activity was associated with fractions containing classic aryl hydrocarbon receptor agonists, such as PCBs, but the mid-polarity fractions containing PAHs and most polar fractions yielded significant dioxinlike activity. Overall, most of the in vitro bioassay responses appear to have been caused by unidentified and/or undetectable compounds associated with Lake Shihwa sediment. Keywords-AlkylphenolsSediment In vitro bioassay Polycyclic aromatic hydrocarbons Polychlorinated biphenyls Estrogen-and dioxinlike potency of sediment contaminants

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Section: Methodsmentioning

confidence: 99%

Alkylphenols, polycyclic aromatic hydrocarbons, and organochlorines in sediment from Lake Shihwa, Korea: Instrumental and bioanalytical characterization

Khim

Villeneuve

Kannan

et al. 1999

Enviro Toxic and Chemistry

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“…Before luciferase was quantified, cytotoxicity was analyzed using a Live/Dead 1 assay (Molecular Probes), which does not interfere with luciferase activity [33,34]. The cytotoxicity assay was performed as described previously by the manufacturer [35] and is based on two reagents, calcein AM and ethidium homodimer-1.…”

Section: Mda-kb2 Assay Protocolmentioning

confidence: 99%

Characterization of the androgen‐sensitive MDA‐kb2 cell line for assessing complex environmental mixtures

Blake

Martinović

Gray

et al. 2010

Enviro Toxic and Chemistry

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Synthetic and natural steroidal androgens and estrogens and many other non-steroidal endocrine-active compounds commonly occur as complex mixtures in aquatic environments. It is important to understand the potential interactive effects of these mixtures to properly assess their risk. Estrogen receptor agonists exhibit additivity in mixtures when tested in vivo and in vitro. Little is known, however, concerning possible mixture interactions of androgen receptor agonists. In these studies we used the MDA-kb2 cell line, a human breast cancer cell line with endogenous androgen receptors and a stably transfected luciferase reporter gene construct to quantify the androgenic activity of seven natural and synthetic androgens: 17beta-trenbolone, dihydrotestosterone, methyltestosterone, testosterone, trendione, 17alpha-trenbolone, and androstenedione. We tested combinations of these androgens and compared the observed activity to expected androgenic activity based on a concentration addition model. Our analyses support the hypothesis that androgen receptor agonists cause additive responses in a mixture. Binary mixtures of 17beta-trenbolone with 17beta-estradiol or triclocarban (an anti-microbial found in the environment) were also tested. 17beta-Estradiol induced androgenic activity, but only at concentrations 600-fold greater than those found in the environment. Triclocarban enhanced the activity of 17beta-trenbolone. Additionally, three anti-androgens were each paired with three androgens of varying potencies. The relative potencies of the antagonists were a vinclozolin metabolite (M2) > procymidone > prochloraz regardless of the androgen used. The results of our studies demonstrate the potential utility of the androgen-responsive MDA-kb2 cell line for quantifying the activity of mixtures of endocrine-active chemicals in complex wastes such as municipal effluents and feedlot discharges.

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“…If the dose-response curves of CYP1A1 protein and catalytic activity differ significantly, other methods of TCDD-EQ determination should be used. Sanderson et al (1996) suggest the use of a recombinant H4IIE cell line transfected with a luciferase reporter gene under control of dioxinresponsive elements (DREs) because luciferase activity is insensitive to substrate inhibition in hepatoma cells (Sanderson et al, 1996;Richter et al, 1997). It is important, however, that the stability of the reporter gene is demonstrated over time before this assay can be used routinely like the wild-type H4IIE cell bioassay.…”

Section: A Accuracymentioning

confidence: 99%

The H4IIE Cell Bioassay as an Indicator of Dioxin-like Chemicals in Wildlife and the Environment

Whyte

Schmitt

Tillitt

2004

Critical Reviews in Toxicology

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ABSTRACT:The H4IIE cell bioassay has proven utility as a screening tool for planar halogenated hydrocarbons (PHHs) and structurally similar chemicals accumulated in organisms from the wild. This bioassay has additional applications in hazard assessment of PHH exposed populations. In this review, the toxicological principles, current protocols, performance criteria, and field applications for the assay are described. The H4IIE cell bioassay has several advantages over the analytical measurement of PHHs in environmental samples, but conclusions from studies can be strengthened when both bioassay and analytical chemistry data are presented together. Often, the bioassay results concur with biological effects in organisms and support direct measures of PHHs. For biomonitoring purposes and prioritization of PHH-contaminated environments, the H4IIE bioassay may be faster and less expensive than analytical measurements. The H4IIE cell bioassay can be used in combination with other biomarkers such as in vivo measurements of CYP1A1 induction to help pinpoint the sources and identities of dioxin-like chemicals. The number of studies that measure H4IIE-derived TCDD-EQs continues to increase, resulting in subtle improvements over time. Further experiments are required to determine if TCDD-EQs derived from mammalian cells are adequate predictors of toxicity to non-mammalian species. The H4IIE cell bioassay has been used in over 300 published studies, and its combination of speed, simplicity, and ability to integrate the effects of complex contaminant mixtures makes it a valuable addition to hazard assessment and biomonitoring studies.

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An in vitro rainbow trout cell bioassay for aryl hydrocarbon receptor‐mediated toxins

Cited by 59 publications

References 46 publications

Alkylphenols, polycyclic aromatic hydrocarbons, and organochlorines in sediment from Lake Shihwa, Korea: Instrumental and bioanalytical characterization

Alkylphenols, polycyclic aromatic hydrocarbons, and organochlorines in sediment from Lake Shihwa, Korea: Instrumental and bioanalytical characterization

Characterization of the androgen‐sensitive MDA‐kb2 cell line for assessing complex environmental mixtures

The H4IIE Cell Bioassay as an Indicator of Dioxin-like Chemicals in Wildlife and the Environment

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