Wu Xinyan, Zahari Muhammad S, Renuse Santosh, Jacob Harrys K C, Sakamuri Sruthi, Singal Mukul, Gabrielson Edward, Sukumar Saraswati, Pandey Akhilesh
a McKusick-Nathans Institute of Genetic Medicine and Department of Biological Chemistry ; Johns Hopkins University School of Medicine ; Baltimore , MD USA.
Cancer Biol Ther. 2014;15(12):1593-9. doi: 10.4161/15384047.2014.961886.
Tissue microarrays (TMAs) have become an invaluable tool in cancer research to evaluate expression and subcellular localization of proteins in cells and tissues. As the catalogs of candidate biomarkers and therapeutic targets become more extensive, there is a need to characterize and validate these targets and biomarkers in cell lines as a primary biological system in research laboratories. Thus, cell microarrays (CMAs) are useful as a high-throughput screening tool. Here, we constructed a CMA containing 32 publicly available immortalized breast cell lines with the goal of creating a method to rapidly screen for antigens of interest in breast cancer research in a relatively easy, rapid and cost-effective manner. As proof of concept, we performed immunocytochemical staining of the HER2 receptor, as the status of this protein is relevant to breast cancer and has previously been reported for these cell lines. We observed a complete concordance of our staining with the published status of HER2 in these cell lines. In addition, we examined the expression of CD44, epithelial markers EpCAM and E-cadherin and tyrosine phosphoproteins. The labeling of these proteins correlates with the known biology of the cell lines. Our results demonstrate the utility of our method to screen for potential biomarkers and therapeutic targets in breast cancer and we suggest that CMAs be used as a general approach in breast cancer research.
组织微阵列(TMAs)已成为癌症研究中一种极具价值的工具,用于评估细胞和组织中蛋白质的表达及亚细胞定位。随着候选生物标志物和治疗靶点的目录日益丰富,有必要在作为研究实验室主要生物系统的细胞系中对这些靶点和生物标志物进行表征和验证。因此,细胞微阵列(CMAs)作为一种高通量筛选工具很有用。在此,我们构建了一个包含32种公开可用的永生化乳腺癌细胞系的细胞微阵列,目的是以相对简便、快速且经济高效的方式创建一种在乳腺癌研究中快速筛选感兴趣抗原的方法。作为概念验证,我们对HER2受体进行了免疫细胞化学染色,因为该蛋白的状态与乳腺癌相关,且此前已针对这些细胞系进行过报道。我们观察到我们的染色结果与这些细胞系中HER2的已发表状态完全一致。此外,我们检测了CD44、上皮标志物EpCAM和E-钙黏蛋白以及酪氨酸磷酸化蛋白的表达。这些蛋白质的标记与细胞系的已知生物学特性相关。我们的结果证明了我们的方法在筛选乳腺癌潜在生物标志物和治疗靶点方面的实用性,并且我们建议将细胞微阵列用作乳腺癌研究中的一种通用方法。