Global gene expression analysis of estrogen receptor transcription factor cross talk in breast cancer: Identification of estrogen-induced/activator protein-1-dependent genes
Global gene expression analysis of estrogen receptor transcription factor cross talk in breast cancer: Identification of estrogen-induced/activator protein-1-dependent genes
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DOI：
10.1210/me.2004-0267
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发表时间：
2005-02-01
影响因子：
--
通讯作者：
Brown, PH
中科院分区：
文献类型：
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作者：
DeNardo, DG;Kim, HT;Brown, PH
越来越多的文献支持雌激素通过非经典途径影响基因表达，其中雌激素受体(ER)调节其他转录因子的活性，如激活蛋白(AP)-1、特异性蛋白(Sp-1)或核因子-kappaB(NFkappaB)。我们假设许多雌激素诱导的基因依赖于AP-1的表达，并且这些基因可以用基因组策略来识别。利用表达可诱导的cJun显性阴性的细胞，我们研究了在AP-1正常或被阻断的条件下雌激素对基因的诱导。我们发现AP-1依赖的基因的表达被cJun显性负性抑制，并且AP-1的阻断不影响ERα的表达或雌激素诱导的雌激素反应元件的活性。然后利用微阵列方法，我们鉴定了20个新的雌激素诱导/AP-1依赖的基因。这些雌激素诱导/AP-1依赖的基因在其启动子中含有更高频率的共同AP-1位点，并且与雌激素诱导的表达不受AP-1阻断影响的基因相比，它们对AP-1刺激剂十四酰佛波醇乙酸酯的敏感性增加。我们还展示了雌激素和AP-1依赖的ER、类固醇受体共激活因子-1和p300通过染色质免疫沉淀对这些基因的启动子的募集。这些研究表明，微阵列可以用于反向遗传学方法来预测受多个转录因子调控的大量基因的功能启动子结构。
There is a growing body of literature supporting estrogen's ability to affect gene expression through a nonclassical pathway, in which estrogen receptor (ER) modulates the activity of other transcription factors such as activator protein (AP)-1, specificity protein (Sp-1), or nuclear factor-kappaB (NFkappaB). We hypothesized that many estrogen-induced genes are dependent on AP-1 for their expression and that these genes can be identified using genomic strategies. Using cells expressing an inducible cJun dominant negative, we studied the estrogen induction of genes under conditions in which AP-1 was normal or blocked. We show that the expression of AP-1-dependent genes was inhibited by the cJun dominant negative and that AP-1 blockade does not affect mRNA ERalpha expression or estrogen induction of estrogen-responsive element activity. Using a microarray approach, we then identified 20 new estrogen-induced/AP-1-dependent genes. These estrogen-induced/AP-1-dependent genes contain a higher frequency of consensus AP-1 sites in their promoters and have increased sensitivity to the AP-1 stimulant tetradecanoyl phorbol acetate when compared with estrogen-induced genes whose expression was not affected by AP-1 blockade. We also show estrogen and AP-1-dependent recruitment of ER, steroid receptor coactivator-1, and p300 to the promoter of these genes by chromatin immunoprecipitation. These studies demonstrate that microarrays can be used in a reverse genetics approach to predict the functional promoter structure of large numbers of genes that are regulated by multiple transcription factors.