Quantitative proteomic analysis to discover potential diagnostic markers and therapeutic targets in human renal cell carcinoma
Quantitative proteomic analysis to discover potential diagnostic markers and therapeutic targets in human renal cell carcinoma
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DOI：
10.1002/pmic.200700619
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发表时间：
2008-08-01
期刊：
影响因子：
3.4
通讯作者：
Okumura, Katsuhiko
中科院分区：
文献类型：
--
作者：
Okamura, Noboru;Masuda, Taro;Okumura, Katsuhiko
肾细胞癌(RCC)对化疗和放射治疗相对耐药。药物开发的最新进展为肾癌的治疗提供了新的药物，但效果仍然很小。此外，迫切需要确定肾癌的诊断标记物。在这份报告中，为了发现潜在的诊断标记和治疗靶点，我们使用2-硝基苯磺基(NBS)试剂对肾癌样本进行了定量蛋白质组学分析。从肾癌和邻近的正常组织中提取蛋白质，从患者的手术中获得，并用含有6个C-12或C-13的NBS试剂标记。紧随其后的是胰酶消化和标记多肽的浓缩。样品经MALDI-TOF MS分析，MS/MS鉴定出差异6 Da的NBS标记的多肽，其中34个蛋白在超过60%的患者中上调，其中一些是已知的，一些是新的。用Western blotting和实时定量RT-PCR鉴定了部分蛋白质的同源性。结果表明，基于NBS的定量蛋白质组学分析有助于发现肾癌的诊断标志和治疗靶点。
Renal cell carcinoma (RCC) is relatively resistant to chemotherapy and radiotherapy. Recent advances in drug development are providing novel agents for the treatment of RCC, but the effects are still minimal. In addition, there is an urgent need to identify diagnostic markers for RCC. In this report, to discover potential diagnostic markers and therapeutic targets, we subjected RCC samples to a quantitative proteomic analysis utilizing 2-nitrobenzenesulfenyl (NBS) reagent. Proteins were extracted from RCC and adjacent normal tissue, obtained surgically from patients, and labeled with NBS reagent containing six C-12 or C-13. This was followed by trypsin digestion and the enrichment of labeled peptides. Samples were then subjected to analysis by MALDI-TOF MS. NBS-labeled peptides with a 6 Da difference were identified by MS/MS. Thirty-four proteins were upregulated in more than 60% of the patients of which some were previously known, and some were novel. The identity of a few proteins was confirmed by Western blotting and quantitative real time RT-PCR. The results suggest that NBS-based quantitative proteomic analysis is useful for discovering diagnostic markers and therapeutic targets for RCC.