Comprehensive analysis of the effect of phytoestrogen, daidzein, on a testicular cell line, using mRNA and protein expression profile

Department of Genomic Drug Discovery Science, Graduate School of Pharmaceutical Sciences, Kyoto University, Yoshida-Shimoadachicho, Sakyo-ku, Kyoto 606-8501, Japan.
Food and Chemical Toxicology (Impact Factor: 2.61). 05/2005; 43(4):529-35. DOI: 10.1016/j.fct.2004.12.006
Source: PubMed

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.

  • [Show abstract] [Hide abstract]
    ABSTRACT: Soybean products contain high concentrations of phytoestrogens. Isoflavone is the major phytoestrogen in soy and has been shown to display estrogen-like activities. It is well known that the delivery of isoflavone components across the placenta and in milk occurs in fetuses and infants the effect of following maternal exposure to isoflavone. We examined the effect of in utero and lactational exposure to isoflavone on the development and fertility of mouse offspring over two generations (F1 and F2). Pregnant mice were given daily subcutaneous injections of isoflavone (5, 50, 100 mg/kg/day) during the gestational and lactational stages until 20 days after delivery. At postnatal day (PND) 42, the F1 pups were sacrificed, and their body masses and organs (liver, kidney, testis, epididymis, ovary and uterus) were weighed. The other F1 pups were mated with non-treated mice or F1 heterosexual mice to estimate F1 fertility and development of F2 pups. The body weights of F1 pups increased at dose of 5 mg/kg/day. The epididymis weights of F1 males increased at doses of 5 and 100 mg/kg/day. No significant differences were found in the uterus and ovary weights of F1 females in all treatments. In the F2 males, the body weights of the 50 mg/kg/day group were higher than those of the control group. The epididymis weights of the 5 and 100 mg/kg/day groups were higher than those of the control group. In the F2 females, the body weights of the 5 and 100 mg/kg/day groups and the ovary weights of the 5 and 50 mg/kg/day groups were higher than those of the control group. There was no significant difference in pregnancy rate or numbers of live or dead fetuses in any treatment for the F1 and F2 offspring. These results suggest that gestational and lactational exposure to isoflavone presumably affect the body weights and some reproductive organ weights of F1 and F2 offspring, but are not seriously harmful to potential fertility.
    Journal of Mammalian Ova Research 01/2009; DOI:10.1274/0916-7625-25.4.272
  • [Show abstract] [Hide abstract]
    ABSTRACT: The effect of long-term exposure of goldfish to dietary genistein and diadzein on the concentrations of plasma sex steroids (testosterone (T), 17β-estradiol (E2)) and the gonadosomatic index (GSI) was assessed. The study was conducted on four groups for a period of 2 years, from the age of 20 weeks to first spawning. Four doses of genistein and diadzein were applied in the feed: genistein: 0 µg/g, diadzein: 0 µg/g (control group); genistein: 24.26 µg/g, diadzein: 21.7µg/g (diet 1); genistein: 51.55 µg/g, diadzein: 46.13 µg/g (diet 2); and genistein: 75.83 µg/g, diadzein: 67.82µg/g (diet 3). Throughout the experiment, there were no significant dose- or time-related effects of genistein and diadzein contents on the T level in both sexes. Furthermore, at the highest genistein and diadzein contents, there was an elevating plasma concentration of E2 at all sampling points (p < 0.05) and a time-related effect occurred (p < 0.05). Although the E2 concentrations in the plasma of female, throughout the experiment, were higher than in males, at the last sampling, the plasma concentrations of E2 reduced among females and became lower than that in males. The effects of isoflavone content were found on GSI of females at the fourth and fifth sampling among the treatments. Isoflavone contents also affect GSI of males at the second, fourth and the last sampling. Our findings suggest that overall genistein and diadzein exposure in early life stages can cause alterations in the reproductive organs and influence sex steroidogenesis.
    Toxicology and Industrial Health 07/2012; 30(2). DOI:10.1177/0748233712452604 · 1.71 Impact Factor
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Epidemiological studies have shown that Asian populations display a lower incidence of hormone-dependant cancers, cardiovascular disease, osteoporosis, and menopausal ailments compared to Western societies. Available data support the proposal that lower incidence is associated with the high dietary consumption of isoflavones, such as genistein. This study used two-dimensional electrophoresis to characterize the effect of genistein on the proteome of an endometrial tumor cell model, namely the Ishikawa cell line. Proteome maps displaying approx 1800 proteins were obtained from cells treated with vehicle or genistein at physiologically attainable concentrations of 0.5, 5, or 50 μM or supra-physiological concentration, 500 μM. The effects of genistein on protein expression were characterized using image analysis software. A total 65 protein spots displayed a significant decrease in expression and 32 proteins displayed a significant increase in expression. Of these protein spots, 29 were randomly selected for characterization by matrix assisted laser desorption/ionization tandem mass spectrometry, yielding 18 different proteins. This type of analysis enabled the characterization of a wide range of cellular proteins and allowed for the identification of functional and biochemical pathways that may be regulated or affected by genistein, including cellular transcription, cell proliferation, stress response, or modulation of oncogenic pathways.
    Clinical Proteomics 07/2006; 2(3):153-167. DOI:10.1007/BF02752498