Darby Ian A, Vuillier-Devillers Karine, Pinault Emilie, Sarrazy Vincent, Lepreux Sébastien, Balabaud Charles, Bioulac-Sage Paulette, Desmoulière Alexis
Cancer Microenviron. 2010 Jun 26;4(1):73-91. doi: 10.1007/s12307-010-0047-2.
Cholangiocarcinoma is an adenocarcinoma of the liver which has increased in incidence over the last thirty years to reach similar levels to other liver cancers. Diagnosis of this disease is usually late and prognosis is poor, therefore it is of great importance to identify novel candidate markers and potential early indicators of this disease as well as molecules that may be potential therapeutic targets. We have used a proteomic approach to identify differentially expressed proteins in peripheral cholangiocarcinoma cases and compared expression with paired non-tumoral liver tissue from the same patients. Two-dimensional fluorescence difference gel electrophoresis after labeling of the proteins with cyanines 3 and 5 was used to identify differentially expressed proteins. Overall, of the approximately 2,400 protein spots visualised in each gel, 172 protein spots showed significant differences in expression level between tumoral and non-tumoral tissue with p < 0.01. Of these, 100 spots corresponding to 138 different proteins were identified by mass spectrometry: 70 proteins were over-expressed whereas 68 proteins were under-expressed in tumoral samples compared to non-tumoral samples. Among the over-expressed proteins, immunohistochemistry studies confirmed an increased expression of 14-3-3 protein in tumoral cells while α-smooth muscle actin and periostin were shown to be overexpressed in the stromal myofibroblasts surrounding tumoral cells. α-Smooth muscle actin is a marker of myofibroblast differentiation and has been found to be a prognostic indicator in colon cancer while periostin may also have a role in cell adhesion, proliferation and migration and has been identified in other cancers. This underlines the role of stromal components in cancer progression and their interest for developing new diagnostic or therapeutic tools.
胆管癌是一种肝癌，在过去三十年中发病率不断上升，已达到与其他肝癌相似的水平。这种疾病的诊断通常较晚，预后较差，因此，识别这种疾病的新型候选标志物、潜在早期指标以及可能成为潜在治疗靶点的分子具有重要意义。我们采用蛋白质组学方法来识别外周胆管癌病例中差异表达的蛋白质，并将其表达与同一患者配对的非肿瘤肝脏组织进行比较。用花菁3和5标记蛋白质后，采用二维荧光差异凝胶电泳来识别差异表达的蛋白质。总体而言，在每块凝胶中可视化的约2400个蛋白质斑点中，有172个蛋白质斑点在肿瘤组织和非肿瘤组织之间的表达水平存在显著差异（p < 0.01）。其中，通过质谱鉴定出与138种不同蛋白质相对应的100个斑点：与非肿瘤样本相比，肿瘤样本中有70种蛋白质过表达，68种蛋白质低表达。在过表达的蛋白质中，免疫组织化学研究证实肿瘤细胞中14-3-3蛋白的表达增加，而α-平滑肌肌动蛋白和骨膜蛋白在肿瘤细胞周围的基质肌成纤维细胞中过表达。α-平滑肌肌动蛋白是肌成纤维细胞分化的标志物，已被发现是结肠癌的预后指标，而骨膜蛋白可能也在细胞黏附、增殖和迁移中起作用，并且已在其他癌症中得到确认。这突出了基质成分在癌症进展中的作用以及它们在开发新的诊断或治疗工具方面的价值。
