Profiling and verification of gene expression patterns in normal and malignant human prostate tissues by cDNA microarray analysis
Profiling and verification of gene expression patterns in normal and malignant human prostate tissues by cDNA microarray analysis
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DOI：
10.1038/sj.neo.7900126
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发表时间：
2001-01-01
期刊：
影响因子：
4.8
通讯作者：
Macoska, JA
中科院分区：
文献类型：
--
作者：
Chaib, H;Cockrell, EK;Macoska, JA
cDNA 微阵列技术可以对几乎任何细胞材料的基因表达模式进行“分析”。在这项研究中，我们应用 cDNA 微阵列技术来分析与人类前列腺肿瘤发生相关的基因表达变化。检查从正常和恶性前列腺组织中制备的 RNA 中 588 个人类基因的表达水平。使用四种不同的数据标准化方法。其中，ACTB 表达标准化被证明是最严格的技术，产生虚假结果的可能性最小。在对 ACTB 表达进行标准化后，通过阵列分析检查的 588 个基因中有 15 个（2.6%）在恶性前列腺组织中与正常前列腺组织相比差异表达达 2 倍或更多。前列腺组织中 15 个基因中的 8 个（TGF beta3、TGFBR3、IGFII、IGFBP2、VEGF、FGF7、ERBB3、MYC）的表达模式先前已报道，但其中 7 个基因（MLH1、CYP1B1、RFC4、EPHB3、MGST1、BTEB2、MLP）的表达模式在此首次报道。这些基因描述了至少四种可能在人类前列腺肿瘤发生中被破坏的代谢和信号传导途径。逆转录酶聚合酶链式反应 (RT-PCR) 和参考 ACTB 表达水平定量的 Northern 印迹分析分别验证了通过阵列分析观察到的 14/15 和 8/8 基因的基因表达水平趋势。然而，RT-PCR 和 Northern blot 分析仅分别准确验证了 6/15 (40%) 和 7/8 (88%) 所检查基因的表达水平“倍数”差异，表明需要更好地验证基于阵列的技术揭示的基因表达的定量差异。
cDNA microarray technology allows the "profiling" of gene expression patterns for virtually any cellular material. In this study, we applied cDNA microarray technology to profile changes in gene expression associated with human prostate tumorigenesis. RNA prepared from normal and malignant prostate tissue was examined for the expression levels of 588 human genes. Four different methods for data normalization were utilized. Of these, normalization to ACTB expression proved to be the most rigorous technique with the least probability of producing spurious results. After normalization to ACTB expression, 15 of 588 (2.6%) genes examined by array analysis were differentially expressed by a factor of 2x or more in malignant compared to normal prostate tissues. The expression patterns for 8 of 15 genes have been reported previously in prostate tissues (TGF beta3, TGFBR3, IGFII, IGFBP2, VEGF, FGF7, ERBB3, MYC), but those of seven genes are reported here for the first time (MLH1, CYP1B1, RFC4, EPHB3, MGST1, BTEB2, MLP). These genes describe at least four metabolic and signaling pathways likely disrupted in human prostate tumorigenesis. Reverse transcriptase polymerase chain reaction (RT-PCR) and Northern blot analyses quantitated with reference to ACTB expression levels verified the trends in gene expression levels observed by array analysis for 14/15 and 8/8 genes, respectively. However, RT-PCR and Northern blot analyses accurately verified the "fold" differences in expression levels for only 6/15 (40%) and 7/8 (88%) of genes examined, respectively, demonstrating the need to better validate quantitative differences in gene expression revealed by array-based techniques.