通过可分离抗体微阵列染色对乳腺癌细胞的蛋白质亚细胞定位进行分析。
Protein subcellular localization profiling of breast cancer cells by dissociable antibody microarray staining.
机构信息
Department of Biological Sciences, Louisiana State University, Baton Rouge, LA 70803, USA.
出版信息
Proteomics. 2010 Apr;10(8):1536-44. doi: 10.1002/pmic.200900585.
We have developed dissociable antibody microarray (DAMA) staining technology that provides a new approach to the global analysis of protein subcellular localization (SCL) in fixed cells. We have developed and optimized this technology for protein SCL profiling, generated ChipView, a program for management and analysis of molecular image database, and utilized the technique to identify proteins with unique SCL in breast cancer cell lines. We compared the SCL profiles of 325 proteins among nine different breast cell lines, and have identified one protein, Cyclin B1, with distinctively different SCLs between normal and cancer cell lines. With classic individual immunostaining, Cyclin B1 was confirmed to localize to the cytoplasm of seven breast cancer cell lines and in both cytoplasm and nuclei of two normal breast cell lines, and to have higher expression levels in the cancer cell lines tested.
我们开发了可分离抗体微阵列（DAMA）染色技术，为固定细胞中蛋白质亚细胞定位（SCL）的全局分析提供了一种新方法。我们已经开发并优化了这项技术用于蛋白质 SCL 分析，生成了 ChipView，这是一个用于管理和分析分子图像数据库的程序，并利用该技术鉴定了乳腺癌细胞系中具有独特 SCL 的蛋白质。我们比较了 9 种不同乳腺癌细胞系中 325 种蛋白质的 SCL 谱，鉴定出一种蛋白质 Cyclin B1，其在正常和癌细胞系之间具有明显不同的 SCL。通过经典的免疫组化染色，Cyclin B1 被证实定位于 7 种乳腺癌细胞系的细胞质中，两种正常乳腺细胞系的细胞质和细胞核中，并且在测试的癌细胞系中表达水平更高。
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