The application of a three-step proteome analysis for identification of new biomarkers of pancreatic cancer.
The application of a three-step proteome analysis for identification of new biomarkers of pancreatic cancer.
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DOI：
10.1155/2011/628787
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发表时间：
2011
期刊：
影响因子：
--
通讯作者：
Nomura F
中科院分区：
文献类型：
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作者：
Abulaizi M;Tomonaga T;Satoh M;Sogawa K;Matsushita K;Kodera Y;Obul J;Takano S;Yoshitomi H;Miyazaki M;Nomura F
我们通过三步血清蛋白质组分析寻找胰腺癌新的肿瘤标志物。十二血清丰富的蛋白质耗尽使用免疫亲和柱，然后通过反相高效液相色谱法分级分离。通过二维凝胶电泳分离每个级分中的蛋白质。然后用考马斯亮蓝染色。通过LC-MS/MS鉴定了癌症和正常对照之间表达水平显著不同的蛋白质点。在从胰腺癌患者获得的血清样品中，102个点上调，84个点下调，并且通过质谱鉴定了58个蛋白质。这些候选蛋白进行了验证，使用蛋白质印迹分析和酶联免疫吸附试验（ELISA）。作为这些验证过程的结果，我们可以确认胰腺癌患者的载脂蛋白A-IV、维生素D结合蛋白、血浆视黄醇结合蛋白4和四连接素的血清水平显著降低。
We searched for novel tumor markers of pancreatic cancer by three-step serum proteome analysis. Twelve serum abundant proteins were depleted using immunoaffinity columns followed by fractionation by reverse-phase high-performance liquid chromatography. Proteins in each fraction were separated by two-dimensional gel electrophoresis. Then the gel was stained by Coomassie Brilliant Blue. Protein spots in which the expression levels were significantly different between cancer and normal control were identified by LC-MS/MS. One hundred and two spots were upregulated, and 84 spots were downregulated in serum samples obtained from patients with pancreatic cancers, and 58 proteins were identified by mass spectrometry. These candidate proteins were validated using western blot analysis and enzyme-linked immunosorbent assay (ELISA). As a result of these validation process, we could confirm that the serum levels of apolipoprotein A-IV, vitamin D-binding protein, plasma retinol-binding protein 4, and tetranectin were significantly decreased in patients with pancreatic cancer.