Cyclin D1 Determines Estrogen Signaling in the Mammary Gland In Vivo
Cyclin D1 Determines Estrogen Signaling in the Mammary Gland In Vivo
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DOI：
10.1210/me.2013-1065
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发表时间：
2013-09-01
影响因子：
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通讯作者：
Pestell, Richard G.
中科院分区：
文献类型：
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作者：
Casimiro, Mathew C.;Wang, Chenguang;Pestell, Richard G.
CCND1 基因在癌症中经常过度表达，它编码磷酸化视网膜母细胞瘤蛋白的全酶的调节亚基。尽管已知细胞周期蛋白 D1 使用异源报告系统调节雌激素受体 (ER)α 反式激活，但细胞周期蛋白 D1 对雌激素依赖性信号传导的体内生物学意义以及细胞周期蛋白 D1 参与的分子机制尚未阐明。在此，对 17 只经 β-雌二醇处理、删除了 Ccnd1 基因的去势处女小鼠进行的全基因组表达谱表明，细胞周期蛋白 D1 决定体内 88% 雌激素响应基因的雌激素依赖性基因表达。此外，17β-雌二醇刺激的细胞周期蛋白 D1 小干扰 RNA 处理的 MCF7 细胞的表达谱显示，细胞周期蛋白 D1 是体外雌激素介导的基因表达所必需的。全基因组染色质免疫沉淀-Seq 分析揭示了与以细胞周期蛋白 D1 依赖性方式受雌激素调节的基因相关的细胞周期蛋白 D1-DNA 结合形式。体内鉴定的细胞周期蛋白 D1 依赖性雌激素信号通路高度富集细胞外膜相关生长因子受体（表皮生长因子受体、ErbB3 和 EphB3）及其配体（双调蛋白，由 AREG 基因编码）和基质金属蛋白酶。 AREG 蛋白是表皮生长因子受体促进细胞增殖的关键配体，由细胞周期蛋白 D1 通过 AREG 启动子诱导。染色质免疫沉淀分析表明细胞周期蛋白 D1 被募集至 Areg 基因的乳腺癌 1 (Brca1)/ER α 结合位点。细胞周期蛋白 D1 基因缺失证明了细胞周期蛋白 D1 在体内组装乳腺癌 1 相关多蛋白复合物中雌激素依赖性扩增的需要。目前的研究定义了体内控制生长因子受体和配体表达的雌激素依赖性信号模块对细胞周期蛋白 D1 的需求，并揭示了细胞周期蛋白 D1 在 ER α 靶基因启动子上的非典型功能。细胞周期蛋白 D1 在生长因子基因的共同顺式元件处介导 ER α 和生长因子信号传导的汇聚。
The CCND1 gene, which is frequently overexpressed in cancers, encodes the regulatory subunit of a holoenzyme that phosphorylates the retinoblastoma protein. Although it is known that cyclin D1 regulates estrogen receptor (ER)alpha transactivation using heterologous reporter systems, the in vivo biological significance of cyclin D1 to estrogen-dependent signaling, and the molecular mechanisms by which cyclin D1 is involved, are yet to be elucidated. Herein, genome-wide expression profiling conducted of 17 beta-estradiol-treated castrated virgin mice deleted of the Ccnd1 gene demonstrated that cyclin D1 determines estrogen-dependent gene expression for 88% of estrogen-responsive genes in vivo. In addition, expression profiling of 17 beta-estradiol-stimulated cyclin D1 small interfering RNA treated MCF7 cells shows cyclin D1 is required for estrogen-mediated gene expression in vitro. Genome-wide chromatin immunoprecipitation-Seq analysis revealed a cyclin D1-DNA bound form associated with genes that were regulated by estrogen in a cyclin D1-dependent manner. The cyclin D1-dependent estrogen signaling pathways identified in vivo were highly enriched for extracellular membrane-associated growth factor receptors (epidermal growth factor receptor, ErbB3, and EphB3) and their ligands (amphiregulin, encoded by AREG gene), and matrix metalloproteinase. The AREG protein, a pivotal ligand for epidermal growth factor receptors to promote cellular proliferation, was induced by cyclin D1 via the AREG promoter. Chromatin immunoprecipitation analysis demonstrated the recruitment of cyclin D1 to the breast cancer 1 (Brca1)/ER alpha binding site of the Areg gene. Cyclin D1 genetic deletion demonstrated the in vivo requirement for cyclin D1 in assembling the estrogen-dependent amplified in breast cancer 1-associated multiprotein complex. The current studies define a requirement for cyclin D1 in estrogen-dependent signaling modules governing growth factor receptor and ligand expression in vivo and reveal a noncanonical function of cyclin D1 at ER alpha target gene promoters. Cyclin D1 mediates the convergence of ER alpha and growth factor signaling at a common cis-element of growth factor genes.