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  • Antiandrogenic mechanisms of pesticides in human LNCaP prostate and H295R adrenocortical carcinoma cells.

Antiandrogenic mechanisms of pesticides in human LNCaP prostate and H295R adrenocortical carcinoma cells.

Toxicological sciences : an official journal of the Society of Toxicology (2014-10-18)
Christina N Robitaille, Patricia Rivest, J Thomas Sanderson
ABSTRACT

Several pesticides suspected or known to have endocrine disrupting effects were screened for pro- or antiandrogenic properties by determining their effects on proliferation, prostatic-specific antigen (PSA) secretion and androgen receptor (AR) expression, and AR phosphorylation in androgen-dependent LNCaP human prostate cancer cells, as well as on the expression and catalytic activity of the enzyme CYP17 in H295R human adrenocortical carcinoma cells, an in vitro model of steroidogenesis. Effects on SRD5A gene expression were determined in both cell lines. Benomyl, vinclozolin, and prochloraz, but not atrazine, concentration dependently (1-30 μM) decreased dihydrotestosterone (DHT)-stimulated proliferation of LNCaP cells. All pesticides except atrazine decreased DHT-stimulated PSA secretion, AR nuclear accumulation, and AR phosphorylation on serines 81 and 213 in LNCaP cells. Benomyl and prochloraz, but not vinclozolin or atrazine, decreased levels of CYP17 gene and protein expression, as well as catalytic activity in H295R cells. In the case of prochloraz, some of these effects corresponded with cytotoxicity. H295R cells expressed AR protein and SRD5A1, but not SRD5A2 transcripts. SRD5A1 gene expression in H295R cells was increased by 10 nM DHT, whereas in LNCaP cells significant induction was observed by 0.1 nM DHT. AR protein expression in H295R cells was not increased by DHT. Vinclozolin decreased DHT-induced SRD5A1 gene expression in LNCaP, but not H295R cells, indicating a functional difference of AR between the cell lines. In conclusion, pesticides may exert antiandrogenic effects through several mechanisms that are cell type-specific, including AR antagonism and down-regulation or catalytic inhibition of androgen biosynthetic enzymes, such as CYP17 and SRD5A1.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Sodium selenite, BioReagent, suitable for cell culture, ≥98%
Sigma-Aldrich
Linoleic acid, technical, 58-74% (GC)
Supelco
Vinclozolin, PESTANAL®, analytical standard
Supelco
Linoleic acid, analytical standard
Sigma-Aldrich
Methyl 1-(butylcarbamoyl)-2-benzimidazolecarbamate, 95%
Sigma-Aldrich
Sodium selenite, γ-irradiated, lyophilized powder, BioXtra, suitable for cell culture
Sigma-Aldrich
Sodium selenite, 99%
Sigma-Aldrich
Linoleic acid, ≥98%
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Linoleic acid, liquid, BioReagent, suitable for cell culture
Sigma-Aldrich
Sodium selenite, anhydrous, ≥90.0% (RT)
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Anti-phospho-Androgen Receptor (Ser81) Antibody, Upstate®, from rabbit
Supelco
Atrazine, certified reference material, TraceCERT®, Manufactured by: Sigma-Aldrich Production GmbH, Switzerland
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L-Glutamine, Pharmaceutical Secondary Standard; Certified Reference Material
Supelco
Atrazine, analytical standard
Supelco
L-Glutamine, certified reference material, TraceCERT®, Manufactured by: Sigma-Aldrich Production GmbH, Switzerland
Sigma-Aldrich
L-Glutamine
Sigma-Aldrich
Dimethyl sulfoxide, anhydrous, ≥99.9%
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8-Octanoyloxypyrene-1,3,6-trisulfonic acid trisodium salt, suitable for fluorescence, ≥90% (HPCE)
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Phenol Red, ACS reagent
Sigma-Aldrich
Dimethyl sulfoxide, sterile-filtered, BioPerformance Certified, meets EP, USP testing specifications, suitable for hybridoma
Supelco
Atrazine, PESTANAL®, analytical standard
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Benomyl, PESTANAL®, analytical standard
Sigma-Aldrich
L-Glutamine, BioUltra, ≥99.5% (NT)
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Dimethyl sulfoxide, analytical standard
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Prochloraz, PESTANAL®, analytical standard
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Forskolin, from Coleus forskohlii, ≥98% (HPLC), powder
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Phenol Red, powder, BioReagent, suitable for cell culture
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Dimethyl sulfoxide, Molecular Biology
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Dimethyl sulfoxide, Hybri-Max, sterile-filtered, BioReagent, suitable for hybridoma, ≥99.7%
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Dimethyl sulfoxide, meets EP testing specifications, meets USP testing specifications