Guedj Nathalie, Dargere Delphine, Degos Françoise, Janneau Jean Louis, Vidaud Dominique, Belghiti Jacques, Bedossa Pierre, Paradis Valerie
CNRS UMR 8149, Faculté de Pharmacie, Paris V, Paris, France.
Lab Invest. 2006 Sep;86(9):951-8. doi: 10.1038/labinvest.3700450. Epub 2006 Jul 17.
Cirrhosis is a heterogeneous tissue composed of polyclonal regenerative and monoclonal neoplastic, potentially malignant nodules from which hepatocellular carcinoma (HCC) might develop. The aim of this study was to investigate proteomic profile changes associated with clonal expansion of cirrhotic nodules and malignant transformation of monoclonal nodules. Seventy-one cirrhotic nodules from 10 female patients with six HCC were dissected from liver surgical specimen by laser capture microdissection. Clonal status of each nodule was assessed by the study of X-chromosome inactivation pattern using the human androgen receptor. Protein profiles were determined by surface-enhanced laser desorption ionisation-time-of-flight technology using Q10 arrays (Cyphergen ProteinChip). Molecular weight of differentially expressed protein peaks was assessed. An average of 50 protein peaks was obtained for each nodule's profile. Comparison of protein profiles in polyclonal (n=45) and monoclonal cirrhotic nodules (n=26) identified three differentially expressed protein peaks (10,092, 54,025 and 62,133 Da). All were upregulated in monoclonal nodules. Twelve peaks were differentially expressed between monoclonal nodules and HCC with nine proteins upregulated in cancer samples. This study confirms that proteome analysis can be achieved from a limited number of microdissected cells, and provides further insight into the process of clonal expansion and malignant transformation of cirrhotic nodules.
肝硬化是一种异质性组织，由多克隆再生结节和单克隆肿瘤性、潜在恶性结节组成，肝细胞癌（HCC）可能由此发展而来。本研究的目的是调查与肝硬化结节的克隆性扩增和单克隆结节的恶性转化相关的蛋白质组学谱变化。通过激光捕获显微切割技术从10例女性患者的肝脏手术标本中分离出71个肝硬化结节，其中6例患有HCC。使用人类雄激素受体通过研究X染色体失活模式来评估每个结节的克隆状态。使用Q10阵列（Cyphergen ProteinChip）通过表面增强激光解吸电离飞行时间技术测定蛋白质谱。评估差异表达蛋白质峰的分子量。每个结节的谱平均获得50个蛋白质峰。多克隆（n = 45）和单克隆肝硬化结节（n = 26）的蛋白质谱比较确定了三个差异表达的蛋白质峰（10,092、54,025和62,133 Da）。所有这些在单克隆结节中均上调。单克隆结节与HCC之间有12个峰差异表达，其中9种蛋白质在癌症样本中上调。本研究证实可以从有限数量的显微切割细胞中进行蛋白质组分析，并为肝硬化结节的克隆性扩增和恶性转化过程提供了进一步的见解。
