Molecular & Cellular Proteomics, 2011, 10(1):657-672.

Quantitative Proteomics and Transcriptomics Addressing the Estrogen Receptor Subtype-mediated Effects in T47D Breast Cancer Cells Exposed to the Phytoestrogen Genistein

Ana M Sotoca, et al

Toxicology section, Wageningen University, Tuinlaan 5, 6703 HE Wageningen, The Netherlands; et al

The present study addresses, by transcriptomics and quantitative stable isotope labeling by amino acids in cell culture (SILAC)-based proteomics, the estrogen receptor α (ERα) and β (ERβ)-mediated effects on gene and protein expression in T47D breast cancer cells exposed to the phytoestrogen genistein. Using the T47D human breast cancer cell line with tetracycline-dependent ERβ expression (T47D-ERβ), the effect of a varying intracellular ERα/ERβ ratio on genistein-induced gene and protein expression was characterized. Results obtained reveal that in ERα-expressing T47D-ERβ cells with inhibited ERβ expression genistein induces transcriptomics and proteomics signatures pointing at rapid cell growth and migration by dynamic activation of cytoskeleton remodeling. The data reveal an interplay between integrins, focal adhesion kinase, CDC42, and actin cytoskeleton signaling cascades, occurring upon genistein treatment, in the T47D-ERβ breast cancer cells with low levels of ERα and no expression of ERβ. In addition, data from our study indicate that ERβ-mediated gene and protein expression counteracts ERα-mediated effects because in T47D-ERβ cells expressing ERβ and exposed to genistein transcriptomics and proteomics signatures pointing at a clear down-regulation of cell growth and induction of cell cycle arrest and apoptosis were demonstrated. These results suggest that ERβ decreases cell motility and metastatic potential as well as cell survival of the breast cancer cell line. It is concluded that the effects of genistein on proteomics and transcriptomics end points in the T47D-ERβ cell model are comparable with those reported previously for estradiol with the ultimate estrogenic effect being dependent on the relative affinity for both receptors and on the receptor phenotype (ERα/ERβ ratio) in the cells or tissue of interest.

美国《分子与细胞蛋白质组学》，2011年1月

当 T47D 乳腺癌细胞暴露于雌激素金雀异黄酮时用定量的蛋白质组学和转录组学来定位雌激素受体的亚型介导作用

安娜 M 萨特克等

荷兰瓦赫宁根大学毒理学院

目前的研究表明，通过基于细胞培养 (SILAC)蛋白质组学的转录组学和定量氨基酸稳定同位素标记，雌激素受体 α （ERα） 和 β （ERβ）-介导对暴露于雌激素金雀异黄酮的T47D 乳腺癌细胞的基因和蛋白表达的影响。利用四环素依赖型ERβ 表达的T47D 人体乳腺癌细胞（T47D-ERβ），表征不同的细胞内 ERα/ERβ对染料木黄酮诱导的基因和蛋白表达的影响。得到的结果表明在 ERα 得到表达而ERβ 表达被抑制的T47D-ERβ 细胞中，金雀异黄素通过细胞骨架重构的动态激活诱使转录组学和蛋白质组学标记快速的细胞生长和迁移。数据揭示了当用 金雀异黄素处理ERα水平 低并且 ERβ 没有表达 的T47D-ERβ 乳腺癌细胞时，整联蛋白、 粘着斑激酶、 CDC42 和肌动蛋白细胞骨架信号级联之间的相互作用。此外，我们的研究数据表明，ERβ 介导的基因和蛋白表达阻碍了ERα 介导的效应，因为在 T47D-e β 细胞表达 ERβ 并暴露于金雀异黄素时，转录组学和蛋白质组学标记的细胞生长和诱导细胞周期阻滞和凋亡明确下调被证明。这些结果表明，ERβ 降低了乳腺癌细胞的细胞活性和侵袭转移潜能以及存活率。可以得出结论，金雀异黄素对 T47D-ERβ 细胞模型中的蛋白质组学和转录组学结束点的影响能够媲美以前报告的雌二醇最终的雌激素作用，这种影响依赖于被关注细胞或组织中两种受体的相对亲和性以及受体表型 （ERα/ERβ之 比）。