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EVALUATION OF THE EFFECTS OF ENVIRONMENTAL COMPOUNDS ON STEROID HORMONE PRODUCTION IN H295R CELLS
Citation:
BUCKALEW, A. R., J. W. LASKEY, J. M. GOLDMAN, E. B. HIGLEY, M. HECKER, AND R. L. COOPER. EVALUATION OF THE EFFECTS OF ENVIRONMENTAL COMPOUNDS ON STEROID HORMONE PRODUCTION IN H295R CELLS. Presented at Triangle Consortium for Reproductive Biology, Chapel Hill, NC, January 27, 2007.
Description:
H295R cells constitute a pluripotent cell line that has retained the enzymatic ability to produce steroids along the entire steroidogenic pathway, including C19 androgens and C18 estrogens. For this reason, they have been a valued research tool, and have been employed in an ever-increasing number of studies. The US EPA is interested in the utility of these cells as an in vitro screen for environmental compounds which may have direct effects on steroidogenesis. This study examined the effects of selected chemicals on the production of progesterone, androstenedione, testosterone, estradiol, estrone and corticosterone by these cells in vitro. After a 48h incubation period, the concentration of each hormone was readily detectable in the medium. Using a 24-well plate format with 200,000 cells per well, it was possible to measure by RIA concurrently up to 4 - 5 hormones per well in a single incubation. Exposure of the cells to chemicals known to modify steroidogenesis affected concentrations of the appropriate hormones as expected. For example, forskolin (which increases cyclic AMP activity) induced a dose-dependent increase of each hormone examined. In contrast, incubation with aminoglutethimide (which inhibits multiple P450s) caused a dose-dependent decrease in the production of several hormones. As part of this work, cells were also examined for any potential effects of compound application on cell viability. These data support the usefulness of this approach as a screen for toxicant effects on steroid hormone production and can also provide valuable insight into sites of impact along the steroidogenic pathway.