Raimondo Francesca, Salemi Claudia, Chinello Clizia, Fumagalli Daniela, Morosi Lavinia, Rocco Francesco, Ferrero Stefano, Perego Roberto, Bianchi Cristina, Sarto Cecilia, Pitto Marina, Brambilla Paolo, Magni Fulvio
Department of Experimental Medicine, Univ. of Milano-Bicocca, Via Cadore 48, 20052 Monza, Italy.
Mol Biosyst. 2012 Apr;8(4):1040-51. doi: 10.1039/c2mb05390j. Epub 2012 Feb 8.
Renal cell carcinoma (RCC), the most common neoplasm affecting the adult kidney, is characterised by heterogeneity of histological subtypes, drug resistance, and absence of molecular markers. Two-dimensional difference gel electrophoresis (2-D DIGE) technology in combination with mass spectrometry (MS) was applied to detect differentially expressed proteins in 20 pairs of RCC tissues and matched adjacent normal kidney cortex (ANK), in order to search for RCC markers. After gel analysis by DeCyder 6.5 and EDA software, differentially expressed protein spots were excised from Deep Purple stained preparative 2DE gel. A total of 100 proteins were identified by MS out of 2500 spots, 23 and 77 of these were, respectively, over- and down-expressed in RCC. The Principal Component Analysis applied to gels and protein spots exactly separated the two sample classes in two groups: RCC and ANK. Moreover, some spots, including ANXA2, PPIA, FABP7 and LEG1, resulted highly differential. The DIGE data were also confirmed by immunoblotting analysis for these proteins. In conclusion, we suggest that applying 2-D DIGE to RCC may provide the basis for a better molecular characterization and for the discovery of candidate biomarkers.
肾细胞癌（RCC）是影响成人肾脏的最常见肿瘤，其特征在于组织学亚型的异质性、耐药性以及缺乏分子标志物。二维差异凝胶电泳（2-D DIGE）技术与质谱（MS）相结合，用于检测20对RCC组织和匹配的相邻正常肾皮质（ANK）中差异表达的蛋白质，以寻找RCC标志物。通过DeCyder 6.5和EDA软件进行凝胶分析后，从深紫色染色的制备性2DE凝胶中切下差异表达的蛋白质斑点。在2500个斑点中，通过MS共鉴定出100种蛋白质，其中23种和77种分别在RCC中上调和下调表达。应用于凝胶和蛋白质斑点的主成分分析将两个样本类别准确地分为两组：RCC和ANK。此外，一些斑点，包括膜联蛋白A2（ANXA2）、亲环素A（PPIA）、脂肪酸结合蛋白7（FABP7）和富含亮氨酸重复序列1（LEG1），差异非常显著。这些蛋白质的免疫印迹分析也证实了DIGE数据。总之，我们建议将2-D DIGE应用于RCC可能为更好的分子特征分析和发现候选生物标志物提供基础。
