Differential Proteomic Analysis of Cancer Stem Cell Properties in Hepatocellular Carcinomas by Isobaric Tag Labeling and Mass Spectrometry

• Published in: Journal of proteome research Vol. 12; no. 8; pp. 3573 - 3585
• Main Authors: Ko, Ching-Huai, Cheng, Chieh-Fang, Lai, Chin-Pen, Tzu, Te-Hui, Chiu, Chih-Wei, Lin, Mei-Wei, Wu, Si-Yuan, Sun, Chung-Yuan, Tseng, Hsiang-Wen, Wang, Chun-Chung, Kuo, Zong-Keng, Wang, Ling-Mei, Chen, Sung-Fang
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 02.08.2013
• Subjects: Biomarkers, Tumor - genetics; Biomarkers, Tumor - metabolism; Carcinoma, Hepatocellular - genetics; Carcinoma, Hepatocellular - metabolism; Carcinoma, Hepatocellular - pathology; Cell Line, Tumor; Cell Transformation, Neoplastic - genetics; Cell Transformation, Neoplastic - metabolism; Cell Transformation, Neoplastic - pathology; Drug Resistance, Neoplasm - genetics; Gene Expression; Gene Expression Profiling; Humans; Liver Neoplasms - genetics; Liver Neoplasms - metabolism; Liver Neoplasms - pathology; Mass Spectrometry; Molecular Sequence Annotation; Neoplastic Stem Cells - metabolism; Neoplastic Stem Cells - pathology; Proteome - genetics; Proteome - metabolism; S100 Proteins - genetics; S100 Proteins - metabolism; Staining and Labeling - methods; Vimentin - genetics; Vimentin - metabolism; isobaric tags for relative and absolute quantitation; cancer stem cells; hepatocellular carcinoma; proteomic analysis
• ISSN: 1535-3893; 1535-3907
• DOI: 10.1021/pr4004294

Malignant tumors are relatively resistant to treatment due to their heterogeneous nature, drug resistance, and tendency for metastasis. Recent studies suggest that a subpopulation of cancer cells is responsible for the malignant outcomes. These cells are considered as cancer stem cells (CSC). Although a number of molecules have been identified in different cancer cells as markers for cancer stem cells, no promising markers are currently available for hepatocellular carcinoma cells. In this study, two clones of Hep3B cell lines were functionally characterized as control or CSC-like cells, based on properties including spheroid formation, drug resistance, and tumor initiation. Furthermore, their protein expression profiles were investigated by isobaric tags for relative and absolute quantitation (iTRAQ), and a total of 1,127 proteins were identified and quantified from the combined fractions; 50 proteins exhibited at least 2-fold differences between these two clones. These 50 proteins were analyzed by GeneGo and were found to be associated with liver neoplasms, hepatocellular carcinoma (HCC), and liver diseases. They were also components of metabolic pathways, immune responses, and cytoskeleton remodeling. Among these proteins, the expressions of S100P, S100A14, and vimentin were verified in several HCC cell lines, and their expressions were correlated with tumorigenicity in HCC cell lines. The functional significance of vimentin and S100A14 were also investigated and verified.