Proteomic analysis of differentially expressed proteins in hepatocellular carcinoma developed in patients with chronic viral hepatitis C
Proteomic analysis of differentially expressed proteins in hepatocellular carcinoma developed in patients with chronic viral hepatitis C
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DOI：
10.1002/pmic.200401194
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发表时间：
2005-09-01
期刊：
影响因子：
3.4
通讯作者：
Rosenbaum, J
中科院分区：
文献类型：
--
作者：
Blanc, JF;Lalanne, C;Rosenbaum, J
肝细胞癌（HCC）是慢性病毒性丙型肝炎的主要并发症。HCC的治疗仍然令人失望。因此，识别新的 HCC 标志物对于疾病的早期检测以及肿瘤特异性蛋白作为潜在的治疗靶点具有重要意义。我们使用蛋白质组学方法来鉴定参与 HCC 发展的新蛋白质。通过二维电泳（2-DE）分析了四例由慢性病毒性丙型肝炎发展而来的 HCC 病例，并将结果与配对的相邻非肿瘤肝组织的结果进行比较。对于 MS 指纹图谱，直接从凝胶上切下 HCC 和非肿瘤肝脏之间具有差异强度的蛋白质斑点，并进行处理以进行 MALDI-MS 和 nano-LC-ESI-MS/MS 分析。每个凝胶中观察到大约 850 个斑点。配对样本的比较分析表明，345个蛋白点在非肿瘤组织和肿瘤组织之间的表达水平存在显着差异。在分析的 345 个蛋白质点中，鉴定出对应于 155 种不同蛋白质的 238 个点； 49 个蛋白质上调，106 个蛋白质下调。在这 155 个蛋白质中，有 91 个蛋白质在至少 3 个案例中受到调节。虽然这 91 种蛋白质中的 52 种已被之前的蛋白质组学或转录组学研究描述过，或者已知与肝癌发生有关，但本实验揭示了 39 种新蛋白质在病毒性丙型肝炎发展成的 HCC 中差异表达。通过蛋白质印迹法，在另外 10 例病毒性丙型肝炎患者发展为 HCC 的病例中，证实了两种选定蛋白质（载脂蛋白 E、氯离子细胞内通道 1）的蛋白质积累变化。
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.