Proteomic analysis of primary colon cancer-associated fibroblasts using the SELDI-ProteinChip platform
Proteomic analysis of primary colon cancer-associated fibroblasts using the SELDI-ProteinChip platform
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DOI：
10.1631/jzus.b1100266
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发表时间：
2012-03
期刊：
影响因子：
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通讯作者：
Zhanhuai Wang;K. Ding;Jie-kai Yu;Xiaoting Zhai;Shu-qin Ruan;Shan-wei Wang;Yong-liang Zhu;Shu Zheng;Su-Zhan Zhang
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文献类型：
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作者：
Zhanhuai Wang;K. Ding;Jie-kai Yu;Xiaoting Zhai;Shu-qin Ruan;Shan-wei Wang;Yong-liang Zhu;Shu Zheng;Su-Zhan Zhang
癌症相关成纤维细胞（CAF）是癌症微环境的标志之一。最近的证据表明，CAFs比正常的成纤维细胞更有能力促进癌细胞的生长和迁移。然而，CAFs独特的蛋白质表达尚未完全阐明。本研究旨在通过比较结肠成纤维细胞与正常成纤维细胞之间的差异蛋白表达来研究结肠成纤维细胞的特性。从人结肠癌手术标本和匹配的正常结肠组织中分离原代成纤维细胞。通过免疫染色分析验证细胞群的纯度。提取来自每组细胞的总细胞裂解物和条件培养基，并使用表面增强激光解吸/电离飞行时间质谱（SELDI-TOF-MS）ProteinChip平台进行蛋白质表达分析。两周后，大多数原代细胞显示出典型的成纤维细胞样特征。在6对原代细胞中的4对中，在CAF内检测到α-平滑肌肌动蛋白阳性肌成纤维细胞的比例增加。成纤维细胞活化蛋白在大多数细胞中弱表达，无差异。利用SELDI-TOF-MS ProteinChip平台，在总细胞裂解物中检测到四个蛋白质峰，质荷比（m/z）为1142、3011、4035和4945，在条件培养基中检测到两个蛋白质峰，m/z为1368和1389。在Swiss-Prot数据库中发现的潜在候选蛋白包括形态发生神经肽、FMRFamide相关肽、胰岛素样生长因子II、胸腺素β-4样蛋白3和紧密连接相关蛋白1。使用SELDI-ProteinChip平台，与正常结肠基质成纤维细胞相比，在结肠CAFs中鉴定差异蛋白质表达。结肠CAFs中复杂的蛋白质组变化可能在结肠癌微环境中发挥重要作用。
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. 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. 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. 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.