title: Dose- and Time-Dependent Transcriptional Response of Ishikawa Cells Exposed to Genistein.
abstract: To further define the utility of the Ishikawa cells as a reliable in vitro model to determine the potential estrogenic activity of chemicals of interest, transcriptional changes induced by genistein (GES) in Ishikawa cells at various doses (10 pM, 1 nM, 100 nM, and 10 μM) and time points (8, 24, and 48 h) were identified using a comprehensive microarray approach. Trend analysis indicated that the expression of 5342 unique genes was modified by GES in a dose- and time-dependent manner (P ≤ 0.0001). However, the majority of gene expression changes induced in Ishikawa cells were elicited by the highest dose of GES evaluated (10 μM). The GES' estrogenic activity was identified by comparing the Ishikawa cells' response to GES versus 17 α-ethynyl estradiol (EE, at equipotent doses, ie, 10 μM vs 1 μM, respectively) and was defined by changes in the expression of 284 unique genes elicited by GES and EE in the same direction, although the magnitude of the change for some genes was different. Further, comparing the response of the Ishikawa cells exposed to high doses of GES and EE versus the response of the juvenile rat uterus exposed to EE, we identified 66 unique genes which were up- or down regulated in a similar manner in vivo as well as in vitro Genistein elicits changes in multiple molecular pathways affecting various biological processes particularly associated with cell organization and biogenesis, regulation of translation, cell proliferation, and intracellular transport; processes also affected by estrogen exposure in the uterus of the rat. These results indicate that Ishikawa cells are capable of generating a biologically relevant estrogenic response and offer an in vitro model to assess this mode of action.
authors: *Mason Business Center, The Procter and Gamble Company, Mason, Ohio 45040 naciff.jm@pg.com.
authors: Jorge M
authors: JM
authors: Naciff
authors: *Mason Business Center, The Procter and Gamble Company, Mason, Ohio 45040.
authors: Zubin S
authors: ZS
authors: Khambatta
authors: *Mason Business Center, The Procter and Gamble Company, Mason, Ohio 45040.
authors: Gregory J
authors: GJ
authors: Carr
authors: *Mason Business Center, The Procter and Gamble Company, Mason, Ohio 45040.
authors: Jay P
authors: JP
authors: Tiesman
authors: Department of Cell Biology, Neurobiology, and Anatomy, Vontz Center for Molecular Studies, University of Cincinnati, Cincinnati, Ohio 45267.
authors: David W
authors: DW
authors: Singleton
authors: Department of Cell Biology, Neurobiology, and Anatomy, Vontz Center for Molecular Studies, University of Cincinnati, Cincinnati, Ohio 45267.
authors: Sohaib A
authors: SA
authors: Khan
authors: *Mason Business Center, The Procter and Gamble Company, Mason, Ohio 45040.
authors: George P
authors: GP
authors: Daston
mesh_terms: D000230:Adenocarcinoma / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D000818:Animals; D045744:Cell Line, Tumor; D004305:Dose-Response Relationship, Drug; D016889:Endometrial Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D004958:Estradiol / Q000494:pharmacology; D005260:Female; D020869:Gene Expression Profiling / Q000379:methods; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects*; D019833:Genistein / Q000494:pharmacology*; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis; D048789:Phytoestrogens / Q000494:pharmacology*; D051381:Rats; D014158:Transcription, Genetic / Q000187:drug effects*; D014599:Uterus / Q000187:drug effects / Q000378:metabolism
chemical_list: D048789:Phytoestrogens; D004958:Estradiol; D019833:Genistein
keywords: Ishikawa cells; gene expression profiling; genistein; human uterus.; in vitro; microarrays