RRC ID 1690
Author Ohno K, Araki N, Yanase T, Nawata H, Iida M.
Title A novel nonradioactive method for measuring aromatase activity using a human ovarian granulosa-like tumor cell line and an estrone ELISA.
Journal Toxicol. Sci.
Abstract Aromatase is a key enzyme in steroidogenesis and plays an important role in sexual differentiation, fertility, and carcinogenesis. Importantly, a variety of chemicals in the environment may influence its activity and thereby disrupt endocrine function. In the current studies, we developed a novel nonradioactive method for measuring aromatase activity that uses a specific ELISA for estrone along with KGN human ovary granulosa-like carcinoma cells. This cell line has relatively high aromatase activity, and because it lacks 17alpha-hydroxylase, it secretes little or no androstenedione, 17beta-estradiol, or estrone. Therefore, aromatase activity can be assayed simply by measuring the production of estrone in the culture medium after addition of the substrate, androstenedione. Furthermore, by making a slight change in the commercial ELISA kit and optimizing the experimental conditions, we developed a sensitive aromatase assay that could measure a wide range of estrone concentrations with very low interference by androgens. We used this assay to investigate the effects of 23 chemicals that have been previously reported to affect aromatase activity in vitro. We confirmed that 17 of 23 test chemicals had inhibitory or inducible effects, although the specific effects of some were different than previously reported. In conclusion, we have developed a simple, sensitive, and nonradioactive assay that can be used for large-scale screening of compounds that can disrupt endocrine function by influencing aromatase activity.
Volume 82(2)
Pages 443-50
Published 2004-12
DOI 10.1093/toxsci/kfh292
PII kfh292
PMID 15456920
MeSH Aromatase / analysis* Aromatase / biosynthesis Benomyl / pharmacology Cell Line, Tumor Cell Survival / drug effects Cross Reactions Enzyme Induction / drug effects Enzyme Inhibitors / pharmacology Enzyme-Linked Immunosorbent Assay Estrone / analysis* Estrone / biosynthesis Female Fungicides, Industrial / pharmacology Granulosa Cell Tumor / enzymology* Humans Ovarian Neoplasms / enzymology* Reference Standards Reproducibility of Results Xenobiotics / pharmacology*
IF 3.564
Times Cited 30
INFORMATION Human/Animal Cells