Proteomic analysis of primary colon cancer-associated fibroblasts using the SELDI-ProteinChip platform  

作　　者：Zhan-huai WANG Ke-feng DING Jie-kai YU Xiao-hui ZHAI Shu-qin RUAN Shan-wei WANG Yong-liang ZHU Shu ZHENG Su-zhan ZHANG 

机构地区：[1]Cancer Institute （Key Laboratory of Cancer Prevention and Intervention, China National Ministry of Education, Key Laboratory of Molecular Biology in Medical Sciences, Zhejiang Province, China）, the Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou 310009, China [2]2Department of Oncology, the Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou 310009, China [3]Department of Gastroenterology, the Second Aff~liated Hospital, School of Medicine, Zhejiang University, Hangzhou 310009, China

出　　处：《Journal of Zhejiang University-Science B(Biomedicine & Biotechnology)》2012年第3期159-167,共9页浙江大学学报（英文版）B辑（生物医学与生物技术）

基　　金：supported by the National Natural Science Foundation of China (Nos. 81000892, 81071801, and 30801341);the Research Fund for the Doctoral Program of Higher Education of China (No. 200803351107)

摘　　要：Objective： Cancer-associated fibroblasts （CAFs） are one of the hallmarks of the cancer microenvironment. Recent evidence has indicated that CAFs are more competent in enhancing cancer cell growth and migration than normal fibroblasts. However, the unique protein expression of CAFs has not been fully elucidated. This study aims to investigate the characterizations of colon CAFs by comparing the differential protein expression between CAFs and normal fibroblasts. Methods： Primary fibroblasts were isolated from surgical specimen of human colon cancer and matched normal colonic tissue. Purity of the cell population was verified through immunostain analysis. Total cell lysates and conditioned media from each group of cells were extracted, and protein expression analysis was con- ducted using the surface-enhanced laser desorption/ionization time-of-flight mass spectrometry （SELDI-TOF-MS） ProteinChip platform. Results： Most primary cells showed typical fibroblast-like features after two weeks. Increased proportion of a-smooth muscle actin-positive myofibroblasts was detected within the CAFs in four of the six pairs of primary cells. Fibroblast activation protein was weakly expressed in most cells without differences. Using SELDI-TOF-MS ProteinChip platform, four protein peaks mass over charge ratio （m/z） 1142, 3011, 4035, and 4945 were detected in the total cell lysates, and two protein peaks m/z 1368 and 1389 were detected in the conditioned media. The potential candidate proteins found in the Swiss-Prot database include morphogenetic neuropeptides, FMRFamide-related peptides, insulin-like growth factor II, thymosin 13-4-like protein 3, and tight junction-associated protein 1. Conclusions： Using the SELDI-ProteinChip platform, differential protein expressions were identified in colon CAFs compared with normal colonic stromal fibroblasts. The complex proteomic alternations in colon CAFs may play important roles related to the colon cancer microenvironment.Objective: Cancer-associated fibroblasts (CAFs) are one of the hallmarks of the cancer microenvironment. Recent evidence has indicated that CAFs are more competent in enhancing cancer cell growth and migration than normal fibroblasts. However, the unique protein expression of CAFs has not been fully elucidated. This study aims to investigate the characterizations of colon CAFs by comparing the differential protein expression between CAFs and normal fibroblasts. Methods: Primary fibroblasts were isolated from surgical specimen of human colon cancer and matched normal colonic tissue. Purity of the cell population was verified through immunostain analysis. Total cell lysates and conditioned media from each group of cells were extracted, and protein expression analysis was conducted using the surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF-MS) ProteinChip platform. Results: Most primary cells showed typical fibroblast-like features after two weeks. Increased proportion of α-smooth muscle actin-positive myofibroblasts was detected within the CAFs in four of the six pairs of primary cells. Fibroblast activation protein was weakly expressed in most cells without differences. Using SELDI-TOF-MS ProteinChip platform, four protein peaks mass over charge ratio (m/z) 1142, 3011, 4035, and 4945 were detected in the total cell lysates, and two protein peaks m/z 1368 and 1389 were detected in the conditioned media. The potential candidate proteins found in the Swiss-Prot database include morphogenetic neuropeptides, FMRFamide-related peptides, insulin-like growth factor II, thymosin β-4-like protein 3, and tight junction-associated protein 1. Conclusions: Using the SELDI-ProteinChip platform, differential protein expressions were identified in colon CAFs compared with normal colonic stromal fibroblasts. The complex proteomic alternations in colon CAFs may play important roles related to the colon cancer microenvironment.

关 键 词：Colon cancer Cancer microenvironment Cancer-associated fibroblasts Proteomics Surface-enhancedlaser desorption/ionization time-of-flight mass spectrometry （SELDI-TOF-MS） 

分 类 号：R735.35[医药卫生—肿瘤]