%0 Journal Article
%T The Application of a Three-Step Serum Proteome Analysis for the Discovery and Identification of Novel Biomarkers of Hepatocellular Carcinoma
%A Asako Kimura
%A Kazuyuki Sogawa
%A Mamoru Satoh
%A Yoshio Kodera
%A Osamu Yokosuka
%A Takeshi Tomonaga
%A Fumio Nomura
%J International Journal of Proteomics
%D 2012
%I Hindawi Publishing Corporation
%R 10.1155/2012/623190
%X 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 ( ). 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. 1. Introduction Hepatocellular carcinoma (HCC) is one of the most common cancers in the world and is a leading cause of death in many countries. Chronic infection by hepatitis B virus (HBV) or hepatitis C virus (HCV) and cirrhosis are major risk factors for HCC development [1, 2]. At present, HCC surveillance with tumor markers and imaging studies such as ultrasonography (US), computed tomography (CT), and magnetic resonance imaging (MRI) have been recommended for patients with cirrhosis [3, 4]. These imaging studies are expensive and the ultrasound is highly dependent on the ability of the operator. Therefore, more sensitive and specific serum biomarkers for early detection of HCC are desirable. Serum tumor markers for detecting HCC could be divided into 4 categories: oncofetal and glycoprotein antigens, enzymes and isoenzymes, genes, and cytokines. Alpha-fetoprotein (AFP) and protein induced by vitamin-K absence or antagonist-II (PIVKA-II) also called des-gamma-carboxyprothrombin (DCP) are representative tumor markers for the diagnosis of HCC. The elevated level of AFP is observed in only 50¨C70% of patients with HCC and also frequently in patients with cirrhosis or exacerbations of chronic hepatitis [5], and its sensitivity is low in patients with earlier/small tumors [6¨C8].
%U http://www.hindawi.com/journals/ijpro/2012/623190/