The application of a three-step proteome analysis for identification of new biomarkers of pancreatic cancer.

International journal of proteomics Pub Date : 2011-01-01 Epub Date: 2011-10-17 DOI:10.1155/2011/628787
Mayinuer Abulaizi, Takeshi Tomonaga, Mamoru Satoh, Kazuyuki Sogawa, Kazuyuki Matsushita, Yoshio Kodera, Jurat Obul, Shigetsugu Takano, Hideyuki Yoshitomi, Masaru Miyazaki, Fumio Nomura
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引用次数: 18

Abstract

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.

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Abstract Image

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三步蛋白质组分析在胰腺癌新生物标志物鉴定中的应用。
我们通过三步血清蛋白质组分析寻找新的胰腺癌肿瘤标志物。采用免疫亲和柱分离12种血清富集蛋白,反相高效液相色谱分离。通过双向凝胶电泳分离各组分中的蛋白质。然后用考马斯亮蓝染色。采用LC-MS/MS技术鉴定癌组织与正常对照组表达水平有显著差异的蛋白点。在胰腺癌患者的血清样本中,有102个位点表达上调,84个位点表达下调,并通过质谱法鉴定出58种蛋白。这些候选蛋白通过western blot分析和酶联免疫吸附试验(ELISA)进行验证。通过这些验证过程,我们可以证实胰腺癌患者血清载脂蛋白a - iv、维生素d结合蛋白、血浆视黄醇结合蛋白4和四联素水平显著降低。
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