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首页> 外文期刊>Electrophoresis: The Official Journal of the International Electrophoresis Society >Proteome analysis of human liver tumor tissue by two-dimensional gel electrophoresis and matrix assisted laser desorption/ionization-mass spectrometry for identification of disease-related proteins
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Proteome analysis of human liver tumor tissue by two-dimensional gel electrophoresis and matrix assisted laser desorption/ionization-mass spectrometry for identification of disease-related proteins

机译：二维凝胶电泳和基质辅助激光解吸/电离质谱技术对人肝肿瘤组织进行蛋白质组学分析，以鉴定疾病相关蛋白

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Hepatocellular carcinoma (HCC) is a common malignancy worldwide and is a leading cause of death. To contribute to the development and improvement of molecular markers for diagnostics and prognostics and of therapeutic targets for the disease, we have largely expanded the currently available human liver tissue maps and studied the differential expression of proteins in normal and cancer tissues. Reference two-dimensional electrophoresis (2-DE) maps of human liver tumor tissue include labeled 2-DE images for total homogenate and soluble fraction separated on pH 3-10 gels, and also images for soluble fraction separated on pH 4-7 and pH 6-9 gels for a more detailed map. Proteins were separated in the first dimension by isoelectric focusing on immobilized pH gradient (IPG) strips, and by 7.5-17.5% gradient sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) gels in the second dimension. Protein identification was done by peptide mass fingerprinting with delayed extraction-matrix assisted laser desorption/ionization-time of flight-mass spectrometry (DE-MALDI-TOF-MS). In total, 212 protein spots (1117 spots in pH 4-7 map and 95 spots in pH 6-9) corresponding to 127 different polypeptide chains were identified. In the next step, we analyzed the differential protein expression of liver tumor samples, to find out candidates for liver cancer-associated proteins. Matched pairs of tissues from 11 liver cancer patients were analyzed for their 2-DE profiles. Protein expression was comparatively analyzed by use of image analysis software. Proteins whose expression levels were different by more than three-fold in at least 30% (four) of the patients were further analyzed. Numbers of protein spots overexpressed or underexpressed in tumor tissues as compared with nontumorous regions were 9 and 28, respectively. Among these 37 spots, 1 overexpressed and 15 underexpressed spots, corresponding to 11 proteins, were identified. The physiological significance of the differential expressions is discussed. [References: 41]
机译：肝细胞癌（HCC）是全世界常见的恶性肿瘤，并且是主要的死亡原因。为了有助于发展和改进用于疾病诊断和预后的分子标记以及该疾病的治疗靶标，我们在很大程度上扩展了当前可用的人肝组织图谱，并研究了正常和癌症组织中蛋白质的差异表达。人肝肿瘤组织的参考二维电泳图（2-DE）包括标记的2-DE图像，用于在pH 3-10凝胶上分离的总匀浆和可溶级分，以及用于在pH 4-7和pH上分离的可溶级分的图像6至9块凝胶可获得更详细的图。在第一维中，通过等电聚焦于固定的pH梯度（IPG）条，在第二维中通过7.5-17.5％梯度十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）凝胶分离蛋白质。通过肽质量指纹图谱与延迟提取基质辅助的激光解吸/电离飞行时间质谱（DE-MALDI-TOF-MS）进行蛋白质鉴定。总共鉴定出对应于127条不同多肽链的212个蛋白质斑点（pH 4-7图中的1117个斑点和pH 6-9中的95个点）。在下一步中，我们分析了肝肿瘤样品中差异蛋白的表达，以寻找与肝癌相关蛋白的候选物。分析了来自11位肝癌患者的配对组织的2-DE谱。通过使用图像分析软件比较分析蛋白质表达。进一步分析了至少30％（四位）患者中表达水平相差三倍以上的蛋白质。与非肿瘤区域相比，在肿瘤组织中高表达或低表达的蛋白斑点的数目分别为9和28。在这37个斑点中，鉴定出1个过表达的斑点和15个表达不足的斑点，分别对应11种蛋白质。讨论了差异表达的生理意义。 [参考：41]

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