Tannapfel Andrea, Anhalt Kathrin, Häusermann Philip, Sommerer Florian, Benicke Markus, Uhlmann Dirk, Witzigmann Helmut, Hauss Johann, Wittekind Christian
Institute of Pathology, University of Leipzig, Liebigstrasse 26, 04103 Leipzig, Germany.
J Pathol. 2003 Oct;201(2):238-49. doi: 10.1002/path.1420.
Characterization of the protein profiles expressed by hepatocellular carcinomas (HCCs) may identify the genes involved in hepatocellular carcinogenesis and offers the possibility of elucidating clinical biomarkers. In an effort to discover such proteins and pathways that are deregulated in hepatocellular carcinogenesis, cellular proteomes of matched normal liver cells and carcinoma were analysed by tissue microdissection and protein microarrays. Using protein microarrays made up of 83 different antibodies, it was possible to monitor alterations of the protein levels in HCC and non-neoplastic liver tissue. Further analysis of altered proteins was performed using western blot analysis and tissue microarrays (TMAs) containing 210 HCC specimens and corresponding liver tissue. The protein microarray approach revealed differential expression between HCC and normal liver of 32 of the 83 proteins examined: 21 of these were up-regulated and 11 down-regulated. IGF (insulin growth factor) II, ADAM (a disintegrin and metalloproteases) 9, STAT (signal transducers and activators of transcription) 3, SOCS (suppressors of cytokine signalling) 3, and cyclin D1 were significantly up-regulated and collagen I, SMAD 4, FHIT (fragile histidine triad), and SOCS1 were down-regulated. The differential expression of these proteins was confirmed using western blot analysis and TMAs. Correlation of differentially regulated proteins with clinico-pathological data showed that cyclin D1 and SOCS1 were associated with tumour prognosis in univariate analysis, but not multivariate analysis. These data indicate that the development of an array-based approach for the determination of protein profiles in HCC may facilitate the identification of new proteins associated with carcinogenesis or prognosis.
对肝细胞癌（HCC）所表达的蛋白质谱进行表征，可能会鉴定出参与肝细胞癌发生的基因，并为阐明临床生物标志物提供可能性。为了发现肝细胞癌发生过程中失调的此类蛋白质和信号通路，通过组织显微切割和蛋白质微阵列分析了匹配的正常肝细胞和癌细胞的细胞蛋白质组。使用由83种不同抗体组成的蛋白质微阵列，能够监测HCC和非肿瘤性肝组织中蛋白质水平的变化。使用蛋白质印迹分析和包含210个HCC标本及相应肝组织的组织微阵列（TMA）对改变的蛋白质进行了进一步分析。蛋白质微阵列方法揭示了在所检测的83种蛋白质中，有32种在HCC和正常肝脏之间存在差异表达：其中21种上调，11种下调。胰岛素样生长因子（IGF）II、解整合素和金属蛋白酶（ADAM）9、信号转导子和转录激活子（STAT）3、细胞因子信号转导抑制因子（SOCS）3和细胞周期蛋白D1显著上调，而胶原蛋白I、SMAD 4、脆性组氨酸三联体（FHIT）和SOCS1下调。使用蛋白质印迹分析和TMA证实了这些蛋白质的差异表达。差异调节蛋白质与临床病理数据的相关性表明，在单变量分析中细胞周期蛋白D1和SOCS1与肿瘤预后相关，但在多变量分析中并非如此。这些数据表明，开发一种基于阵列的方法来确定HCC中的蛋白质谱可能有助于鉴定与癌发生或预后相关的新蛋白质。
