Screening of some anti-progestin endocrine disruptors using a recombinant yeast based in vitro bioassay

Chatterjee, Shamba; Kumar, Vikas; Majumder, Chandrajeet B.; Roy, Partha

doi:10.1016/j.tiv.2007.12.006

Cited by 39 publications

(19 citation statements)

References 60 publications

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“…EC 50 for budesonide of about 5 μM in yeast and 2 nM in the GR-CALUX. A lower sensitivity was expected as also the yeast-based assays for the detection of oestrogens, androgens and progestagins have been shown to be less sensitive than their mammalian analogues [29, 30, 33, 42–44]. However, the sensitivity of this yeast glucocorticoid assay is relatively very low.…”

Section: Discussionmentioning

confidence: 99%

“…However, the sensitivity of this yeast glucocorticoid assay is relatively very low. This becomes clear when the EC 50 values for E2, P4, T and Dex of respectively 0.5, 1, 50 and 100.000 nM in yeast-based ER, PR, AR and GR bioassays are compared with their U2-OS-based CALUX analogues, with EC 50 values of respectively 0.02, 0.6, 0.7 and 0.7 nM [29, 30, 33, 42–44]. The relatively very low sensitivity of the yeast glucocorticoid bioassay is most probably due to the absence of the human HSP90 chaperone protein; the present data thus suggest that HSP90 is more needed in yeast to obtain optimal hGRα activity than it is needed to obtain fully active ER, PR and AR.…”

Section: Discussionmentioning

confidence: 99%

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Recombinant cell bioassays for the detection of (gluco)corticosteroids and endocrine-disrupting potencies of several environmental PCB contaminants

et al. 2011

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Sensitive and robust bioassays for glucocorticoids are very useful for the pharmaceutical industry, environmental scientists and veterinary control. Here, a recombinant yeast cell was constructed that expresses the human glucocorticoid receptor alpha and a green fluorescent reporter protein in response to glucocorticoids. Both the receptor construct and the reporter construct were stably integrated into the yeast genome. The correct and specific functioning of this yeast glucocorticoid bioassay was studied by exposures to cortisol and other related compounds and critically compared to a GR-CALUX bioassay based on a human bone cell. Although less sensitive, the new yeast glucocorticoid bioassay showed sensitivity towards all (gluco)corticoids tested, with the following order in relative potencies: budesonide >> corticosterone > dexamethasone > cortisol = betamethasone > prednisolone > aldosterone. Hormone representatives for other hormone nuclear receptors, like 17β-estradiol for the oestrogen receptor, 5α-dihydrotestosterone for the androgen receptor and progesterone for the progesterone receptor, showed no clear agonistic responses, whilst some polychlorinated biphenyls were clearly able to interfere with the GR activity.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Recombinant cell bioassays for the detection of (gluco)corticosteroids and endocrine-disrupting potencies of several environmental PCB contaminants

et al. 2011

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“…DDT, methoxychlor and dieldrin). However, the endocrine activity of numerous commonly used compounds like pharmaceutics, products in industrial wastes, pesticides, water treatment plant effluents has not yet been studied in detail so far [2,5,19,25,26]. Out of various EDCs that have been identified till date, majority of them focused on the identification of (anti)estrogenic chemicals from these sources as a result of which the interaction of these chemicals with many other steroid hormones like androgen, progesterone, glucocorticoids and thyroid were understudied.…”

Section: Discussionmentioning

confidence: 99%

Anti-androgenic endocrine disrupting activities of chlorpyrifos and piperophos

Gunda

Chatterjee

Dabral

et al. 2010

The Journal of Steroid Biochemistry and Molecular Biology

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“…Several molecules are described as anti-progesterone as they are able to bind to the progesterone receptor but cannot induce the binding to the response element and thus prevent a biological response [29]. By neutralizing a proportion of free progesterone receptors, these compounds have a modulating effect on progesterone receptor agonists.…”

Section: Anti-progesterone Effectmentioning

confidence: 99%

“…For progesterone detection, similar assays have been developed using mammalian or yeast cells as host for the production of the progesterone receptor and an enzyme or a fluorescent protein as reporter [29][30][31][32][33][34]. Wiswanath et al [35] reported the development of a transactivation assay using modified S. cerevisiae cells producing the human progesterone receptor with a progesterone response element (PRE) controlling the production of a luciferase gene.…”

Section: Introductionmentioning

confidence: 99%

Development of a recombinant Arxula adeninivorans cell bioassay for the detection of molecules with progesterone activity in wastewater

et al. 2015

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This study describes the development of a bioassay to detect the presence of progesterone and progesterone-like molecules in wastewater samples. The basis of the bioassay is the integration of the human progesterone receptor gene into the yeast Arxula adeninivorans for the constitutive synthesis of the receptor. After incubation, binding of the analyte to the receptor induces the production of a reporter protein. Two reporter proteins were compared for detection parameters such as half-maximal activity (EC50), limit of detection (LoD) and limit of quantification (LoQ). When the extracellular phytase K was used, an EC50 value of 155 ng L(-1) and a LoD of 27 ng L(-1) progesterone were obtained after 4 h incubation, while use of the fluorescent dsRED as the reporter protein, resulted in an EC50 of 320 ng L(-1) and a LoD of 65 ng L(-1) after 20 h incubation. Use of phytase K as the reporter protein offers decreased incubation time and increased sensitivity; however the dsRED reporter system is less labor-intensive. Additionally, the affinity of known agonists and antagonists of the human progesterone receptor was determined. The utility of this bioassay was confirmed by measuring total progesterone equivalent concentration of samples from a wastewater treatment plant. The A. adeninivorans-based transactivation assay was able to measure concentrations of about 311 ng L(-1) in the influent stream but could not detect progesterone activity in effluent. One key feature of the assay is the robustness of A. adeninivorans, which allows sample measurement without any sample preparation.

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Screening of some anti-progestin endocrine disruptors using a recombinant yeast based in vitro bioassay

Cited by 39 publications

References 60 publications

Recombinant cell bioassays for the detection of (gluco)corticosteroids and endocrine-disrupting potencies of several environmental PCB contaminants

Recombinant cell bioassays for the detection of (gluco)corticosteroids and endocrine-disrupting potencies of several environmental PCB contaminants

Anti-androgenic endocrine disrupting activities of chlorpyrifos and piperophos

Development of a recombinant Arxula adeninivorans cell bioassay for the detection of molecules with progesterone activity in wastewater

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