Gene expression profiling reveals novel regulation by bisphenol-A in estrogen receptor-α-positive human cells
Gene expression profiling reveals novel regulation by bisphenol-A in estrogen receptor-α-positive human cells
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DOI：
10.1016/j.envres.2005.05.004
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发表时间：
2006-01-01
影响因子：
8.3
通讯作者：
Khan, SA
中科院分区：
文献类型：
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作者：
Singleton, DW;Feng, YX;Khan, SA
双酚A(BPA)在子宫和乳腺中表现出增殖作用，可能影响子宫内或出生后早期男性和女性生殖道的发育。由于其作为雌激素受体(ER)激动剂的功能，BPA有可能通过调节ER靶基因来干扰正常的内分泌信号。有些基因同时受雌二醇(E2)和双酚A(BPA)的调节，但其中任何一种因子所特有的基因尚未被描述。使用将卵黄蛋白原A2 ere-lacZ报告基因整合到基因组中的酵母菌株，我们发现BPA诱导了报告基因在ERα表达载体转化的菌落中的表达，说明了BPA在染色质背景下的调节作用。此外，瞬时导入子宫内膜癌(Ishikawa)细胞系的报告基因也显示出BPA活性，尽管其活性比E2低100倍。为了比较BPA和E2对全球基因表达的响应，我们使用了稳定表达HA标记Era的MCF-7乳腺癌细胞株的变种。培养细胞用乙醇溶液、E2(10(-8)M)或BPA(10(-6)M)处理3h，然后提取RNA，用人类U95A探针阵列(Affymetrix，Santa Clara，CA，USA)进行微阵列分析。超过300个基因被一种或两种药物改变了两倍或更多，大约一半上调，一半下调。一些与生长发育相关的基因，如HOXC1和C6、Wnt5A、FrizzledTM、转化生长因子β-2和STAT抑制因子2，被发现仅受BPA的影响。我们使用实时定量聚合酶链式反应来验证HOXC6基因的调控，结果显示BPA使HOXC6基因的表达减少了约2.5倍。这些结果揭示了BPA和E2的新作用，提出了关于内分泌干扰物在性发育中的作用的有趣的可能性。(C)2005 Elsevier Inc.保留所有权利。
Bisphenol-A (BPA) shows proliferative actions in uterus and mammary glands and may influence the development of male and female reproductive tracts in utero or during early postnatal life. Because of its ability to function as an estrogen receptor (ER) agonist, BPA has the potential to disrupt normal endocrine signaling through regulation of ER target genes. Some genes are regulated by both estradiol (E2) and BPA, but those exclusive to either agent have not been described. Using a yeast strain incorporating a vitellogenin A2 ERE-LacZ reporter gene into the genome., we found that BPA induced expression of the reporter in colonies transformed with the ER alpha expression plasmid, illustrating BPA-mediated regulation within a chromatin context. Additionally, a reporter gene transiently transfected into the endometrial cancer (Ishikawa) cell line also showed BPA activity, although at 100-fold less potency than E2. To compare global gene expression in response to BPA and E2, we used a variant of the MCF-7 breast cancer cell line stably expressing HA-tagged ERa. Cultures were treated for 3 h with an ethanol vehicle, E2 (10(-8) M), or BPA (10(-6)M), followed by isolation of RNA and microarray analysis with the human U95A probe array (Affymetrix, Santa Clara, CA, USA). More than 300 genes were changed 2-fold or more by either or both agents, with roughly half being up-regulated and half down-regulated. A number of growth- and development-related genes, such as HOXC1 and C6, Wnt5A, Frizzled, TGF beta-2, and STAT inhibitor 2, were found to be affected exclusively by BPA. We used quantitative real-time PCR to verify regulation of the HOXC6 gene, which showed decreased expression of approximately 2.5-fold by BPA. These results reveal novel effects by BPA and E2, raising interesting possibilities regarding the role of endocrine disruptors in sexual development. (c) 2005 Elsevier Inc. All rights reserved.