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Differential display screening for specific gene expression induced by dietary nonsteroidal estrogen
Author(s) -
Hsu JihTay,
Jean TzuChao,
Chan MayAl,
Ying Chingwen
Publication year - 1999
Publication title -
molecular reproduction and development
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.745
H-Index - 105
eISSN - 1098-2795
pISSN - 1040-452X
DOI - 10.1002/(sici)1098-2795(199902)52:2<141::aid-mrd4>3.0.co;2-v
Subject(s) - daidzein , biology , differential display , complementary dna , microbiology and biotechnology , gene expression , cell growth , gene , estrogen , messenger rna , northern blot , endocrinology , biochemistry , genistein
The dietary phytoestrogen, daidzein, produced a biphasic response in cell proliferation of cultured, estrogen‐responsive human breast carcinoma MCF‐7 cells. Cell growth was stimulated at a daidzein concentration of 0.25 μg/ml whereas the addition of daidzein at concentrations >25 μg/ml significantly inhibited cell growth in a dose‐dependent fashion, resulting in an IC 50 value of 50 μg/ml. Upon exposure to 50 μg/ml of daidzein, cell morphology was severely altered, cell volume decreased, and condensation of the chromosomes was clearly noticeable. To identify genes whose expression were inhibited by daidzein, a differential display reverse transcriptase polymerase chain reaction assay (DD‐RT‐PCR) was performed and the cDNA fragments of several daidzein‐regulated genes were visualized. The sequence of one of the cloned cDNA fragments that showed differential mRNA expression level in response to daidzein at a concentration of 50 μg/ml had a high homology with a cDNA expressed in fetal human brain, EST 06411. Mol. Reprod. Dev. 52:141–148, 1999. © 1999 Wiley‐Liss, Inc.