title: Comprehensive analysis of the effect of phytoestrogen, daidzein, on a testicular cell line, using mRNA and protein expression profile.
abstract: In this study, we examined the effects of exposure to phytoestrogen (daidzein), 17beta-estradiol (E2), diethylstilbestrol (DES) and staurosporin on the TM4 testicular cell line, using comprehensive analysis, such as cDNA microarray and two-dimension polyacrylamide gel electropholesis (2D-PAGE) analysis, and we demonstrated if these toxicogenomic analyses could classify the chemical compounds. First, RNA was extracted from TM4 cells that had been treated with daidzein (80 microM), DES, E2 (40 microM) and stauroporin (100 nM) for 30 min. We performed cDNA microarray analysis, and the expression ratio data thus obtained were then analyzed using hierarchical clustering. This hierarchical clustering showed that daidzein exposure induced a different effect on gene expression change from that of E2, DES and staurosporin. Next, protein extracted from TM4 cells also underwent cDNA microarray analysis for 3 h. We performed 2D-PAGE analysis, and the spot intensity ratio data thus obtained were analyzed using hierarchical clustering. As with cDNA microarray, the hierarchical clustering of protein spot ratios showed that daidzein exposure induced a different effect on gene expression change from that of the other substances. In conclusion, we have demonstrated for the first time that classification of these chemicals can be performed by clustering analysis, using data from cDNA microarray and 2D-PAGE analyses, and that exposure to daidzein induces effects different from those of E2, DES and staurosporin.
authors.affiliation: Department of Genomic Drug Discovery Science, Graduate School of Pharmaceutical Sciences, Kyoto University, Yoshida-Shimoadachicho, Sakyo-ku, Kyoto 606-8501, Japan. adachi@tom.life.h.kyoto-u.ac.jp
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Adachi
authors.forename: Yasushi
authors.initials: Y
authors.lastname: Okuno
authors.forename: Shigeo
authors.initials: S
authors.lastname: Takenaka
authors.forename: Kazuyuki
authors.initials: K
authors.lastname: Matsuda
authors.forename: Naoki
authors.initials: N
authors.lastname: Ohta
authors.forename: Kyoka
authors.initials: K
authors.lastname: Takashima
authors.forename: Koji
authors.initials: K
authors.lastname: Yamazaki
authors.forename: Daisuke
authors.initials: D
authors.lastname: Nishimura
authors.forename: Kazutaka
authors.initials: K
authors.lastname: Miyatake
authors.forename: Chisato
authors.initials: C
authors.lastname: Mori
authors.forename: Gozoh
authors.initials: G
authors.lastname: Tsujimoto
mesh_terms: D002460:Cell Line; D004054:Diethylstilbestrol / Q000494:pharmacology; D015180:Electrophoresis, Gel, Two-Dimensional; D004958:Estradiol / Q000494:pharmacology; D004968:Estrogens, Non-Steroidal / Q000494:pharmacology; D020869:Gene Expression Profiling; D006801:Humans; D007529:Isoflavones / Q000494:pharmacology*; D008297:Male; D020411:Oligonucleotide Array Sequence Analysis; D048789:Phytoestrogens / Q000494:pharmacology*; D012333:RNA, Messenger / Q000032:analysis; D012708:Sertoli Cells; D013737:Testis / Q000166:cytology* / Q000187:drug effects*
chemical_list: D004968:Estrogens, Non-Steroidal; D007529:Isoflavones; D048789:Phytoestrogens; D012333:RNA, Messenger; D004958:Estradiol; C004742:daidzein; D004054:Diethylstilbestrol