Table of Contents
International Journal of Proteomics
Volume 2012, Article ID 623190, 12 pages
Research Article

The Application of a Three-Step Serum Proteome Analysis for the Discovery and Identification of Novel Biomarkers of Hepatocellular Carcinoma

1Department of Molercular Diagnosis, Graduate School of Medicine, Chiba University, Chiba 260-0856, Japan
2Clinical Proteomics Research Center, Chiba University Hospital, Chiba, Japan
3Laboratory of Biomolecular Dynamics, Department of Physics, Kitasato University School of Science, Kanagawa, Japan
4Department of Medicine and Clinical Oncology, Graduate School of Medicine, Chiba University, Chiba 260-0856, Japan
5Laboratory of Proteome Research, National Institute of Biomedical Innovation, Osaka, Japan

Received 7 February 2012; Accepted 5 June 2012

Academic Editor: Dayan B. Goodenowe

Copyright © 2012 Asako Kimura et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.


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 . 0 2 3 ). 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.