Transcriptional signatures of environmentally relevant exposures in normal human mammary epithelial cells: benzo[a]pyrene
Transcriptional signatures of environmentally relevant exposures in normal human mammary epithelial cells: benzo[a]pyrene
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DOI：
10.1016/j.canlet.2004.08.037
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发表时间：
2005-04-28
期刊：
影响因子：
9.7
通讯作者：
Weston, A
中科院分区：
文献类型：
--
作者：
Keshava, C;Whipkey, D;Weston, A
研究了一组原代正常人乳腺上皮细胞株中基因表达的变化，这些细胞株是从不同供体的乳房缩小成形术中获得的健康乳腺组织发育而来，对苯并[a]芘暴露的反应。预计来自不同供体的细胞株所共有的基因表达变化以及个体间的变异都会被观察到。因此，已采取的策略是利用基因阵列技术和少量捐赠者，从生物监测的角度识别潜在的重要变化或有用的变化；然后使用大量组织供体和更便宜的转录检测方法（实时聚合酶链式反应）研究选定的转录反应。在这里，我们报告了四种原代正常人乳腺上皮细胞株的结果，这些细胞株在体外用苯并[a]芘处理6或24小时。使用高密度寡核苷酸阵列（Affymetrix HuGeneFL）监测转录。总 RNA 用于制备标记靶标，这些靶标与含有代表 6800 多个人类基因和表达序列标签的探针的微阵列杂交。使用GeneChip 软件(MAS 5.0)分析基因表达数据。在来自不同供体的人乳腺上皮细胞株中观察到对苯并[a]芘反应的基因表达模式的改变。具体来说，在来自所有四个供体的细胞株中，二恶英诱导型细胞色素 P450 CYP1B1 在暴露于苯并[a]芘 6 和 24 小时后一致被诱导。另外两个相对一致诱导的基因是 ILI beta 和 MMPI。其他代谢基因（CYP1A1、CYP11B2 和 NQO1）以及某些细胞周期控制基因 GOS2 和 AF1Q 也被诱导出现不太一致的变化，而 EGR1 则受到抑制。尽管没有观察到 p53 转录的变化，但使用抗体检测到 p53 蛋白的积累。使用免疫组织化学也观察到 WafI (p21) 的类似积累，这是预期的，因为 p53 是 p21 的转录因子。苯并[a]芘暴露后，基因表达水平和模式均观察到显着的个体间差异。这些研究为分子流行病学研究化学致癌物（特别是多环芳烃）的不同敏感性提供了补充方法。由爱思唯尔爱尔兰有限公司出版
Changes in gene expression in a panel of primary normal human mammary epithelial cell strains, developed from healthy breast tissue obtained at reduction mammoplasty from different donors, in response to benzo[a]pyrene exposure have been investigated. It was expected that both gene expression changes common to cell strains derived from different donors as well as inter-individual variation would be observed. Therefore, the strategy that has been adopted is to identify potentially important changes, or useful changes from a biomonitoring perspective, using gene-array technology and a small number of donors; then investigate selected transcription responses using a large number of tissue donors and a cheaper method of transcript detection (real-time polymerase chain reaction). Here we report results from four primary normal human mammary epithelial cell strains that were treated with benzo[a]pyrene in vitro for either 6 or 24 h. Transcription was monitored using high-density oligonucleoticle arrays (Affymetrix HuGeneFL). Total RNA was used for the preparation of labeled targets that were hybridized to microarrays containing probes representing more than 6800 human genes and expressed sequence tags. Gene expression data were analyzed using the GeneChip (R) software (MAS 5.0). Altered gene expression patterns were observed in response to benzo[a]pyrene in human mammary epithelial cell strains from different donors. Specifically, the dioxin inducible cytochrome P450 CYP1B1 was consistently induced in response to 6 and 24 h exposure to benzo[a]pyrene in cell strains from all four donors. Two other genes that were relatively consistently induced were ILI beta and MMPI. Less consistent changes in other metabolism genes (CYP1A1, CYP11B2, and NQO1) and certain cell cycle control genes GOS2 and AF1Q were also induced, while EGR1 was suppressed. Although no change in p53 transcription was observed, an accumulation of p53 protein was detected using antibodies. A similar accumulation of WafI (p21) was also observed using immunohistochemistry, this was expected sincep53 is p21's transcription factor. Significant inter-individual variations in both the levels and patterns of gene expression were observed, in response to benzo[a]pyrene exposure. These studies provide a complementary approach to molecular epidemiology for the investigation of differential susceptibility to chemical carcinogens, and specifically polycyclic aromatic hydrocarbons. Published by Elsevier Ireland Ltd.