The application of a three-step serum proteome analysis for the discovery and identification of novel biomarkers of hepatocellular carcinoma.
The application of a three-step serum proteome analysis for the discovery and identification of novel biomarkers of hepatocellular carcinoma.
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DOI：
10.1155/2012/623190
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发表时间：
2012
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：
Kimura A;Sogawa K;Satoh M;Kodera Y;Yokosuka O;Tomonaga T;Nomura F
HCC的代表性肿瘤标志物AFP和PIVKA-II在HCC早期诊断的敏感性和特异性方面并不令人满意。为了寻找新的HCC标志物，我们对12例HCC和10例LC患者的血清样本进行了三步蛋白质组分析。作为第一步，将血清样品置于基于抗体的免疫亲和柱系统中，该系统同时去除12种丰富的血清蛋白。然后使用反相HPLC将浓缩的流出物分级。通过SDS-PAGE分离在每个级分中获得的蛋白质。平行分析从HCC和LC患者获得的血清样品，并比较它们的蛋白质表达模式。共发现83条蛋白条带在HCC血清中上调。所有的蛋白条带，其强度在HCC和LC组之间是不同的，被鉴定。其中，clusterin的表达最为显著（P = 0.023）。使用另一组验证的HCC样品通过ELISA证实血清丛生蛋白的过表达。此外，在AFP和PIVKA-II均在其临界值范围内的HCC病例中，40%的血清丛生蛋白升高。这些结果表明，丛生蛋白是一个潜在的新的血清标志物肝癌。
The representative tumor markers for HCC, AFP, and PIVKA-II are not satisfactory in terms of sensitivity and specificity in the early diagnosis of HCC. In search for novel markers for HCC, three-step proteome analyses were carried out in serum samples obtained from 12 patients with HCC and 10 with LC. As a first step, serum samples were subjected to antibody-based immunoaffinity column system that simultaneously removes twelve of abundant serum proteins. The concentrated flow-through was then fractionated using reversed-phase HPLC. Proteins obtained in each fraction were separated by SDS-PAGE. Serum samples obtained from patient with HCC and with LC were analyzed in parallel and their protein expression patterns were compared. A total of 83 protein bands were found to be upregulated in HCC serum. All the protein bands, the intensity of which was different between HCC and LC groups, were identified. Among them, clusterin was most significantly overexpressed (P = 0.023). The overexpression of serum clusterin was confirmed by ELISA using another validation set of HCC samples. Furthermore, serum clusterin was elevated in 40% of HCC cases in which both AFP and PIVKA-II were within their cut-off values. These results suggested that clusterin is a potential novel serum marker for HCC.