Use of two-dimensional gel electrophoresis in predictive toxicology:: Identification of potential early protein biomarkers in chemically induced hepatocarcinogenesis
Use of two-dimensional gel electrophoresis in predictive toxicology:: Identification of potential early protein biomarkers in chemically induced hepatocarcinogenesis
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DOI：
10.1002/pmic.200401067
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发表时间：
2005-05-01
期刊：
影响因子：
3.4
通讯作者：
Kröger, M
中科院分区：
文献类型：
--
作者：
Fella, K;Glückmann, M;Kröger, M
我们目前的方法侧重于识别潜在的早期蛋白质生物标志物特征，这些特征表明暴露于 20 mg/kg 肝脏致癌物 N-亚硝基吗啉 (NNM) 的大鼠的致癌过程。在不同时间点对处理过的肝脏进行研究。因此，在通过质谱鉴定差异表达的蛋白质之前，第一步是通过二维凝胶电泳分离蛋白质。暴露一天后肝脏样本的蛋白质组学分析显示，与 NNM 诱导的细胞应激反应有关的蛋白质（超氧化物歧化酶、热休克蛋白 60、过氧化还原蛋白）显着上调。停止 NNM 18 周后，我们能够在携带恶性转化细胞的大鼠肝脏中鉴定出癌症相关蛋白（Caspase-8 前体、波形蛋白、Rho GDP 解离抑制剂）。其中一些蛋白质在暴露三周后已经解除管制，表明它们作为肝脏致癌性早期预测生物标志物的潜在用途（膜联蛋白 A5、果糖 1,6-二磷酸酶）。由于监管毒理学方法通常包括在啮齿类动物中进行为期两年的研究中的致癌性调查，特别是在肿瘤出现之前检测早期蛋白质生物标志物特征，这表明蛋白质组学方法在大幅减少致癌性测试的时间和成本方面具有巨大潜力。
Our current approach focused on the identification of potential early protein biomarker signatures which are indicative of the carcinogenic processes in rats exposed to 20 mg/kg of the liver carcinogen N-nitrosomorpholine (NNM). Treated liver was investigated at different timepoints. T'herefore, proteins were separated by two-dimensional gel electrophoresis as a first step prior to identification of differentially expressed proteins by mass spectrometry. Proteomic analysis of liver samples after one day of exposure revealed significant upregulation of proteins involved in response to cellular stress induced by NNM (superoxide dismutase, heat shock protein 60, peroxiredoxin). Eighteen weeks after withdrawal of NNM, we were able to identify cancer-related proteins in rat liver bearing malignant, transformed cells (caspase-8 precursor, vimentin, Rho GDP dissociation inhibitor). Some of these proteins were already deregulated after three weeks of exposure indicating their potential usefulness as early predictive biomarkers for liver carcino-genicity (annexin A5, fructose-1,6-bisphosphatase). As regulatory toxicology approaches usually include the investigation of carcinogenicity in two-years studies in rodents, especially the detection of early protein biomarker signatures which precede the appearance of neoplasia, demonstrates the high potential of proteomics approaches to substantially reduce the time and costs of carcinogenicity testing.