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寡核苷酸阵列与长cDNA阵列揭示的差异基因表达模式。

Differential gene expression patterns revealed by oligonucleotide versus long cDNA arrays.

作者信息

Li Jiang, Pankratz Matthew, Johnson Jeffrey A

机构信息

School of Pharmacy, University of Wisconsin, 777 Highland Avenue, Madison, WI 53706, USA.

出版信息

Toxicol Sci. 2002 Oct;69(2):383-90. doi: 10.1093/toxsci/69.2.383.

Abstract

DNA microarrays can be classified into oligonucleotides (Affymetrix) or long cDNAs (IncyteGenomics) based on the arrayed probes. Unfortunately, data are lacking on the comparison of these two popular global screening array systems. The present study was designed to assess the reliability of datasets generated by the two platforms from the same samples. We have already established a model for upregulation of a cluster of antioxidant responsive element (ARE)-driven genes in a human neuroblastoma cell line by treatment with tert-butylhydroquinone (tBHQ) for 8 and 24 h. HuGene FL (Affymetrix), U95 Av2 (Affymetrix), and UniGem V 2.0 (IncyteGenomics) were chosen to do the comparative study on 8- and 24-h samples. The Affymetrix data generated from U95Av2 chips demonstrated that the mRNA of 218 (2.3% of total clones) genes was increased after 8 h of tBHQ treatment. This list included most of the known ARE-driven genes, and nine selected genes showed a high consistency with RT-PCR results. IncyteGenomics called four genes increased and no genes were decreased. These same four genes were also called by the Affymetrix microarray. The sensitivity (fluorescence intensity) and specificity (fold) were very different for selected genes when comparing the two platforms. Cross-hybridization was shown to partially contribute to the discrepancies of the data generated by the two platforms. According to our results, the data generated from oligonucleotide microarrays is more reliable for interrogating changes in gene expression than data from long cDNA microarrays.

摘要

根据点阵排列的探针,DNA微阵列可分为寡核苷酸微阵列(Affymetrix公司)或长cDNA微阵列(IncyteGenomics公司)。遗憾的是,目前尚缺乏关于这两种常用的全基因组筛选阵列系统比较的数据。本研究旨在评估由这两种平台从相同样本中生成的数据集的可靠性。我们已经建立了一个模型,通过用叔丁基对苯二酚(tBHQ)处理人神经母细胞瘤细胞系8小时和24小时,上调一组抗氧化反应元件(ARE)驱动的基因。选择HuGene FL(Affymetrix公司)、U95 Av2(Affymetrix公司)和UniGem V 2.0(IncyteGenomics公司)对8小时和24小时的样本进行比较研究。从U95Av2芯片生成的Affymetrix数据表明,tBHQ处理8小时后,218个基因(占总克隆数的2.3%)的mRNA水平升高。该列表包括了大多数已知的ARE驱动基因,9个选定基因的结果与逆转录聚合酶链反应(RT-PCR)结果高度一致。IncyteGenomics公司检测到4个基因上调,未检测到基因下调。这4个相同的基因也被Affymetrix微阵列检测到。在比较这两种平台时,选定基因的灵敏度(荧光强度)和特异性(倍数)差异很大。交叉杂交被证明是导致这两种平台生成的数据存在差异的部分原因。根据我们的结果,与长cDNA微阵列数据相比,寡核苷酸微阵列生成的数据在检测基因表达变化方面更可靠。

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