Blanc Jean-Frédéric, Lalanne Céline, Plomion Christophe, Schmitter Jean-Marie, Bathany Katell, Gion Jean-Marc, Bioulac-Sage Paulette, Balabaud Charles, Bonneu Marc, Rosenbaum Jean
INSERM, E362, IFR66 Bordeaux, Université Victor Segalen Bordeaux 2, Bordeaux, France.
Proteomics. 2005 Sep;5(14):3778-89. doi: 10.1002/pmic.200401194.
Hepatocellular carcinoma (HCC) is a major complication of chronic viral hepatitis C. Therapy for HCC is still disappointing. It is thus of great importance to identify novel HCC markers for early detection of the disease, and tumor-specific proteins as potential therapeutic targets. We have used a proteomic approach to identify new proteins involved in HCC development. Four cases of HCC developing from chronic viral hepatitis C were analyzed by two-dimensional electrophoresis (2-DE), and results were compared to those of paired adjacent non-tumorous liver tissues. For MS fingerprinting, protein spots with differential intensity between HCC and non-tumorous liver were directly cut out of gels and processed for MALDI-MS and nano-LC-ESI-MS/MS analysis. Approximately 850 spots were visualized in each gel. The comparative analysis of paired samples indicated that 345 protein spots showed significant differences in expression level between non-tumor and tumor tissue. Among the 345 protein spots analyzed, 238 spots corresponding to 155 different proteins were identified; 49 proteins were up-regulated, whereas 106 proteins were down-regulated. Among these 155 proteins, 91 proteins were regulated in at least three cases. Although 52 out of these 91 proteins have been already described by previous proteomic or transcriptomic studies, or are already known to be involved in hepatocarcinogenesis, this experiment revealed 39 new proteins differentially expressed in HCC developing from viral hepatitis C. Variations in protein accumulation were confirmed for two selected proteins (apolipoprotein E, chloride intracellular channel 1) by Western blotting in ten additional cases of HCC developing in patients with viral hepatitis C.
肝细胞癌（HCC）是慢性丙型病毒性肝炎的主要并发症。HCC的治疗效果仍然令人失望。因此，识别用于早期检测该疾病的新型HCC标志物以及作为潜在治疗靶点的肿瘤特异性蛋白质至关重要。我们采用蛋白质组学方法来识别参与HCC发生发展的新蛋白质。通过二维电泳（2-DE）分析了4例由慢性丙型病毒性肝炎发展而来的HCC病例，并将结果与配对的相邻非肿瘤性肝组织进行比较。对于质谱指纹图谱分析，将HCC与非肿瘤性肝组织之间强度有差异的蛋白质点直接从凝胶中切出，进行基质辅助激光解吸电离质谱（MALDI-MS）和纳升液相色谱-电喷雾串联质谱（nano-LC-ESI-MS/MS）分析。每块凝胶中大约可见850个蛋白点。配对样本的比较分析表明，345个蛋白质点在非肿瘤组织和肿瘤组织之间的表达水平存在显著差异。在分析的345个蛋白质点中，鉴定出了对应于155种不同蛋白质的238个蛋白点；49种蛋白质上调，而106种蛋白质下调。在这155种蛋白质中，91种蛋白质在至少3例中受到调控。尽管这91种蛋白质中有52种已被先前的蛋白质组学或转录组学研究报道过，或者已知参与肝癌发生，但本实验揭示了39种在丙型病毒性肝炎相关HCC中差异表达的新蛋白质。通过蛋白质免疫印迹法在另外10例丙型病毒性肝炎患者发生的HCC中对两种选定蛋白质（载脂蛋白E、细胞内氯离子通道1）的蛋白质积累变化进行了确认。
