Publication details
Quantitative assessment of the effects of chemicals on steroidogenic enzymes in the H295R cell line using real-time quantitative PCR
| Authors | |
|---|---|
| Year of publication | 2004 |
| Type | Article in Periodical |
| Magazine / Source | Toxicological sciences |
| MU Faculty or unit | |
| Citation | |
| Field | Genetics and molecular biology |
| Keywords | Steroidogenesis; bioassay; xenoestrogens; screening |
| Description | The potential for a variety of environmental contaminants to disturb endocrine function in wildlife species has been of recent concern. While much current effort is focused on the assessment of effects mediated through steroid hormone receptor based mechanisms, there is potentially a variety of other mechanisms, which could lead to endocrine disruption. Recent studies have demonstrated that a variety of xenobiotics can alter the gene expression or activity of enzymes involved in steroidogenesis. By altering the production of steroidogenic enzymes these chemicals have the potential to alter the delicate steroid balance in organisms. To assess the potential of chemicals to alter steroidogenesis an assay system was developed based on a human cell line that retains the ability to synthesize most of the important steroidogenic enzymes. Methods were developed to measure the expression of 10 genes involved in steroidogenesis by use of Q-RT-PCR. Assay conditions were optimized and expression levels were normalized to the expression of beta-actin. The effects of a variety of model chemicals known to alter steroid metabolism, both inducers and inhibitors, were assessed. Similar expression patterns were observed for chemicals acting through common mechanisms of action. Time-course studies demonstrated distinct time dependent expression profiles for chemicals able to modulate steroid metabolism. The assay, which allows simultaneous analysis of the expression of numerous steroidogenic enzymes, would be useful as a sensitive and rapid integrative screen for the many effects of chemicals on steroidogenesis. |