title: Integrated Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC) and Isobaric Tags for Relative and Absolute Quantitation (iTRAQ) Quantitative Proteomic Analysis Identifies Galectin-1 as a Potential Biomarker for Predicting Sorafenib Resistance in Liver Cancer.
abstract: Sorafenib has become the standard therapy for patients with advanced hepatocellular carcinoma (HCC). Unfortunately, most patients eventually develop acquired resistance. Therefore, it is important to identify potential biomarkers that could predict the efficacy of sorafenib. To identify target proteins associated with the development of sorafenib resistance, we applied stable isotope labelling with amino acids in cell culture (SILAC)-based quantitative proteomic approach to analyze differences in protein expression levels between parental HuH-7 and sorafenib-acquired resistance HuH-7 (HuH-7(R)) cells in vitro, combined with an isobaric tags for relative and absolute quantitation (iTRAQ) quantitative analysis of HuH-7 and HuH-7(R) tumors in vivo. In total, 2,450 quantified proteins were identified in common in SILAC and iTRAQ experiments, with 81 showing increased expression (>2.0-fold) with sorafenib resistance and 75 showing decreased expression (<0.5-fold). In silico analyses of these differentially expressed proteins predicted that 10 proteins were related to cancer with involvements in cell adhesion, migration, and invasion. Knockdown of one of these candidate proteins, galectin-1, decreased cell proliferation and metastasis in HuH-7(R) cells and restored sensitivity to sorafenib. We verified galectin-1 as a predictive marker of sorafenib resistance and a downstream target of the AKT/mTOR/HIF-1α signaling pathway. In addition, increased galectin-1 expression in HCC patients' serum was associated with poor tumor control and low response rate. We also found that a high serum galectin-1 level was an independent factor associated with poor progression-free survival and overall survival. In conclusion, these results suggest that galectin-1 is a possible biomarker for predicting the response of HCC patients to treatment with sorafenib. As such, it may assist in the stratification of HCC and help direct personalized therapy.
authors: From the ‡Graduate Institute of Biochemistry and Molecular Biology.
authors: Chao-Chi
authors: CC
authors: Yeh
authors: §Graduate Institute of Oncology, College of Medicine, ‖Department of Oncology, National Taiwan University Hospital, Taipei, Taiwan.
authors: Chih-Hung
authors: CH
authors: Hsu
authors: §Graduate Institute of Oncology, College of Medicine, ‖Department of Oncology, National Taiwan University Hospital, Taipei, Taiwan.
authors: Yu-Yun
authors: YY
authors: Shao
authors: From the ‡Graduate Institute of Biochemistry and Molecular Biology.
authors: Wen-Ching
authors: WC
authors: Ho
authors: ¶Institute of Biotechnology, National Taiwan University and.
authors: Mong-Hsun
authors: MH
authors: Tsai
authors: From the ‡Graduate Institute of Biochemistry and Molecular Biology.
authors: Wen-Chi
authors: WC
authors: Feng
authors: From the ‡Graduate Institute of Biochemistry and Molecular Biology, chowip@ntu.edu.tw.
authors: Lu-Ping
authors: LP
authors: Chow
mesh_terms: D000596:Amino Acids; D000818:Animals; D000970:Antineoplastic Agents / Q000627:therapeutic use*; D014408:Biomarkers, Tumor / Q000097:blood / Q000235:genetics / Q000378:metabolism*; D006528:Carcinoma, Hepatocellular / Q000097:blood*; D045744:Cell Line, Tumor; D002465:Cell Movement; D049109:Cell Proliferation; D019008:Drug Resistance, Neoplasm / Q000502:physiology; D058750:Epithelial-Mesenchymal Transition; D037483:Galectin 1 / Q000097:blood / Q000235:genetics / Q000378:metabolism*; D055785:Gene Knockdown Techniques; D006801:Humans; D007553:Isotope Labeling; D008113:Liver Neoplasms / Q000097:blood*; D008807:Mice, Inbred BALB C; D009536:Niacinamide / Q000031:analogs & derivatives* / Q000627:therapeutic use; D010671:Phenylurea Compounds / Q000627:therapeutic use*; D060066:Protein Interaction Maps; D040901:Proteomics / Q000379:methods; D000077157:Sorafenib; D016896:Treatment Outcome
chemical_list: D000596:Amino Acids; D000970:Antineoplastic Agents; D014408:Biomarkers, Tumor; D037483:Galectin 1; D010671:Phenylurea Compounds; D009536:Niacinamide; D000077157:Sorafenib