| Using a Customized DNA Microarray for Expression Profiling of the Estrogen-Responsive Genes to Evaluate Estrogen Activity among Natural Estrogens and Industrial Chemicals Shunichi Terasaka,1,2 Yukie Aita,1 Akio Inoue,1,3 Shinichi
Hayashi,3 Michiko Nishigaki,4 Kazuhiko Aoyagi,4 Hiroki
Sasaki,4 Yuko Wada-Kiyama,5 Yasuo Sakuma,5 Shuichi
Akaba,6 Junko Tanaka,7 Hideko Sone,7Junzo Yonemoto,7 Masao Tanji,8 and Ryoiti Kiyama1,8 1Research Institute for Biological Resources and Functions, National
Institute of Advanced Industrial Science and Technology, Tsukuba, Ibaraki,
Japan; 2SciMedia Ltd., Taito-ku, Tokyo, Japan; 3Division
of Endocrinology, Saitama Cancer Center Research Institute, Komuro, Ina, Saitama,
Japan; 4Genetics Division, National Cancer Center Research Institute,
Tsukiji, Chuo-ku, Tokyo, Japan; 5Department of Physiology, Nippon
Medical School, Sendagi, Bunkyo-ku, Tokyo, Japan; 6Inovation Research
Center, Nissui Pharmaceutical Co., Ltd., Sugamo, Toshima-ku, Tokyo, Japan; 7Endocrine
Disruptors & Dioxin Research Project, National Institute for Environmental
Studies, Tsukuba, Japan; 8InfoGenes Co., Ltd., Tsukuba, Ibaraki,
Japan Abstract
We developed a DNA microarray to evaluate the estrogen activity of natural estrogens and industrial chemicals. Using MCF-7 cells, we conducted a comprehensive analysis of estrogen-responsive genes among approximately 20,000 human genes. On the basis of reproducible and reliable responses of the genes to estrogen, we selected 172 genes to be used for developing a customized DNA microarray. Using this DNA microarray, we examined estrogen activity among natural estrogens (17ß-estradiol, estriol, estrone, genistein) , industrial chemicals (diethylstilbestrol, bisphenol A, nonylphenol, methoxychlor) , and dioxin. We obtained results identical to those for other bioassays that are used for detecting estrogen activity. On the basis of statistical correlations analysis, these bioassays have shown more sensitivity for dioxin and methoxychlor. Key words: endocrine disruptor, estrogenicity, expression profile, microarray. Environ Health Perspect 112:773-781 (2004) . doi:10.1289/txg.6753 available via http://dx.doi.org/ [Online 12 February 2004]
Address correspondence to R. Kiyama, Research Institute for Biological Resources and Functions, National Institute of Advanced Industrial Science and Technology, AIST Central 6, 1-1-1 Higashi, Tsukuba, Ibaraki 305-8566, Japan. Telephone: 81 298 61 6189. Fax: 81 298 61 6190. E-mail: kiyama.r@aist.go.jp This study was funded by a grant from the Ministry of the Environment of Japan for preventing public pollution and by a grant for research and development of small businesses from the Ministry of Economy, Trade and Industry of Japan. The authors declare they have no competing financial interests. Received 22 September 2003 ; accepted 12 February 2004.
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