14-3-3 Protein beta/alpha as a urinary biomarker for renal cell carcinoma: proteomic analysis of cyst fluid
14-3-3 Protein beta/alpha as a urinary biomarker for renal cell carcinoma: proteomic analysis of cyst fluid
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DOI：
10.1007/s00216-011-5057-5
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发表时间：
2011-07-01
影响因子：
4.3
通讯作者：
Baba, Shiro
中科院分区：
文献类型：
--
作者：
Minamida, Satoru;Iwamura, Masatsugu;Baba, Shiro
尽管已经分析了来自肾细胞癌（RCC）患者的各种样品，包括组织、细胞、血清和尿液，但是具有诊断价值的生物标志物尚未被鉴定。我们使用蛋白质组学方法分析囊肿相关RCC病例的囊液，以鉴定RCC细胞过度表达和/或分泌的可获得和丰富的蛋白质。囊液中的蛋白质通过反相高效液相色谱和琼脂糖双向凝胶电泳进行分离，并通过串联质谱进行鉴定。我们进行了国家生物技术信息中心搜索和MEDLINE搜索，以预测这些鉴定的蛋白质的功能，并选择肿瘤标志物候选蛋白质。我们的研究导致了被称为14-3-3蛋白β/α的差异调节蛋白的鉴定和选择，该蛋白在囊肿相关RCC的囊液中过表达，但以前与RCC无关。然后，我们通过各种正常和RCC样本（血清，尿液，组织和囊液）的Western印迹法测量其发病率。14-3-3蛋白β/α的表达水平在RCC患者的尿液样品中高于健康志愿者的样品。进行受试者工作特征（ROC）曲线分析以评估这种潜在的生物标志物;这些数据（ROC曲线下面积值为0.8813）表明这种筛选方法的准确度很高。14-3-3蛋白β/α可能是RCC早期诊断的诊断有用的生物标志物。
Although various samples, including tissue, cells, serum, and urine, from patients with renal cell carcinoma (RCC) have been analyzed, biomarkers with diagnostic value have yet to be identified. We used a proteomics approach to analyze cyst fluid in cases of cyst-associated RCC to identify accessible and abundant proteins that are overexpressed and/or secreted by RCC cells. Proteins in the cyst fluid were separated by reverse-phase high-performance liquid chromatography and agarose two-dimensional gel electrophoresis and were identified by tandem mass spectrometry. We conducted a National Center for Biotechnology Information search and a MEDLINE search to predict the function of these identified proteins and to select a tumor-marker candidate protein. Our search resulted in the identification and selection of the differentially regulated protein known as 14-3-3 protein beta/alpha, which was overexpressed in cyst fluid from cyst-associated RCC but has not been previously associated with RCC. We then measured its incidence through Western blotting of various normal and RCC samples (serum, urine, tissue, and cyst fluid). The expression levels of 14-3-3 protein beta/alpha were higher in urine samples from patients with RCC than in samples from healthy volunteers. Receiver operating characteristic (ROC) curve analyses were performed to assess this potential biomarker; these data (area under the ROC curve value was 0.8813) indicate a high degree of accuracy for this screening method. 14-3-3 Protein beta/alpha may be a diagnostically useful biomarker for early diagnosis of RCC.