Unité 785, INSERM, Villejuif, France.
Mol Cell Proteomics. 2010 Sep;9(9):1991-2004. doi: 10.1074/mcp.M110.000026. Epub 2010 May 31.
Obtaining accurate protein profiles from homogeneous cell populations in heterogeneous tissues can enhance the capability to discover protein biomarkers. In this context, methodologies to access specific cellular populations and analyze their proteome with exquisite sensitivity have to be selected. We report here the results of an investigation using a combination of laser microdissection and accurate mass and time tag proteomics. The study was aimed at the precise determination of proteome alterations in intrahepatic cholangiocarcinoma ICC, a markedly heterogeneous tumor. This cancer, which is difficult to diagnose and carries a very poor prognosis, has shown an unexplained increase in incidence over the last few years. Among a pool of 574 identified proteins, we were able to report on altered abundance patterns affecting 39 proteins conforming to a variety of potential tumorigenic pathways. The reliability of the proteomics results was confirmed by Western blot and immunohistochemistry on matched samples. Most of the proteins displaying perturbed abundances had not yet been described in the setting of ICC. These include proteins involved in cell mobility and actin cytoskeleton remodeling, which may participate in the epithelial to mesenchymal transition, a process invoked in migration and invasion of cancer cells. The biological relevance of these findings was explored using a tissue microarray. An increased abundance of vimentin was thus detected in 70% of ICC and none of the controls. These results suggest that vimentin could play a role in the aggressiveness of ICC and provide a basis for the serious outcome of this cancer.
从异质组织中的同质细胞群中获取准确的蛋白质谱，可以提高发现蛋白质生物标志物的能力。在这种情况下，必须选择能够获取特定细胞群并以极高的灵敏度分析其蛋白质组的方法。我们在此报告了使用激光显微切割和精确质量和时间标记蛋白质组学相结合的研究结果。该研究旨在精确确定肝内胆管癌（ICC）的蛋白质组改变，ICC 是一种明显异质的肿瘤。这种癌症难以诊断，预后极差，近年来发病率却呈上升趋势，原因不明。在鉴定出的 574 种蛋白质中，我们能够报告影响 39 种蛋白质丰度变化的模式，这些蛋白质涉及多种潜在的致癌途径。Western blot 和匹配样本的免疫组织化学验证了蛋白质组学结果的可靠性。大多数显示丰度变化的蛋白质在 ICC 中尚未被描述过。这些蛋白质包括参与细胞迁移和肌动蛋白细胞骨架重塑的蛋白质，它们可能参与上皮细胞向间充质转化过程，这是癌细胞迁移和侵袭过程中所涉及的过程。使用组织微阵列探索了这些发现的生物学相关性。在 70%的 ICC 中检测到波形蛋白的丰度增加，而在对照组中则没有。这些结果表明，波形蛋白可能在 ICC 的侵袭性中起作用，并为这种癌症的严重后果提供了依据。
