A functional array for signature of breast cancer

描述（由申请人提供）： 乳腺癌是由遗传和表观遗传变化的积累引起的。基因芯片技术已被广泛用于识别乳腺癌相关基因表达的变化，用于乳腺肿瘤的分类和预后。然而，从cDNA微阵列分析获得的信息不足以准确地表征基因产物的生物学状态。蛋白质功能的癌症相关变化的分析在阐明癌症发生的分子机制方面提供了很多信息。转录因子是调节基因表达的重要细胞蛋白。我们最近开发了蛋白质/DNA阵列技术，能够同时分析多个转录因子的激活。我们建议验证，在第一阶段的研究中，如果这种新的阵列技术可用于确定癌症相关的TF活性模式，通过分析乳腺癌细胞系中的300个转录因子的活动。我们将开发（1）玻璃微阵列来取代目前的膜基阵列和（2）硝基纤维素基旋转柱来取代蛋白质/DNA复合物的琼脂糖凝胶分离。通过这些改进，整个分析过程可以自动化，这将使同时分析大量样品变得更加容易。我们将应用这项技术分析20个乳腺癌细胞系，这些细胞系来自不同阶段的乳腺癌以及一个正常的乳腺癌细胞系，并比较它们之间的转录因子活性模式，以确定乳腺癌转录因子的特征。

DESCRIPTION (provided by applicant): Breast cancer is caused by the accumulation of genetic and epigenetic changes. The cDNA microarray technology has been widely used for identifying cancer-related changes in gene expression for breast tumor classification and prognosis. However, the information obtained from cDNA microarray analysis is not sufficient to accurately characterize the biological state of gene products. The profiling of cancer-related changes in protein functions is much informative in elucidating the molecular mechanism by which cancer occurs. Transcription factors are important cellular proteins, which regulate gene expression. We have recently developed protein/DNA array technology that is capable of profiling the activation of multiple transcription factors simultaneously. We propose to verify, in the phase I study, if this novel array technology can be used to identify cancer-related patterns of TF activity by profiling the activities of 300 transcription factors in breast cancer cell lines. We will develop (1) a glass microarray to replace the current membrane-based array and (2) nitrocellulose-based spin column to replace agarose gel separation of protein/DNA complexes. With the improvements, the entire assay procedures can be automated, which will make it much easier to analyze a lot of samples simultaneously. We will apply this technology to analyze 20 breast cancer cell lines that are derived from different stages of breast cancers as well as one normal breast cell line and to compare the patterns of transcription factor activity among them in order to identify the signature of transcription factors for breast cancers.