US EPA

7351767 

Journal Article 

Endocrine disrupting chemicals - Assaying androgenicity by quantitating the induced expression levels in two different prostate cell lines: Human prostate carcinoma cells 22RV1 and normal primary porcine epithelial cells 

Hartel, A; Pertl, C; Wierer, M; Meyer, HHD 

2004 

1 

Fresenius Environmental Bulletin
ISSN: 1018-4619
EISSN: 1610-2304 

13 

11A 

1129-1138 

WOS:000226019700010 

First systematic data revealed that some "endocrine disruptors" act through mimicking or antagonising the physiological functions of estrogens. Later, endocrine disrupting pathways were described to influence the signal cascade of the androgen receptor (AR). Difenoconazole and fentinacetate (TPTA) are used as fungicides, a third-tetramethrin-as an insecticide. We have already established a gene expression assay for testing chemicals for androgen activity, which is performed in human prostate carcinoma 22RV1 cells growing androgen independently. A second newly established assay is based on normal primary porcine prostate epithelial cells (NPE). We have identified about nine androgen-regulated genes in the prostate, four of which are eligible to determine androgenic action using highly sensitive real-time RT-PCR: AR, differentiation related gene1 (drg1), 5alpha-reductase and cycline D-1. Expression of mRNA for AR and drg1 gene by 22RV1 and NPE cells shows a similar course after treatment with fentinacetate and testosterone. In both assays the upregulation was 2-5 fold for two investigated genes. Based on these results we provide indications that fentinacetate is an endocrine disruptor with strong androgenic effects. Final goal of this work is to establish a sensitive system for differential gene expression by different compounds under study, substance-specific expression patterns and function related analysis of potential androgenic side activity. 

22RV1 cells; normal primary porcine prostate cells; fentinacetate; TPT; gene expression; real-time RT-PCR