Mass Spectrometric Identification of Human Prostate Cancer-derived Proteins in Serum of Xenograft-bearing Mice*
Mass Spectrometric Identification of Human Prostate Cancer-derived Proteins in Serum of Xenograft-bearing Mice*
复制标题
DOI：
10.1074/mcp.m500371-mcp200
复制
发表时间：
2006-09
影响因子：
7
通讯作者：
G. van den Bemd;J. Krijgsveld;T. Luider;A. V. van Rijswijk;J. Demmers;G. Jenster
中科院分区：
文献类型：
--
作者：
G. van den Bemd;J. Krijgsveld;T. Luider;A. V. van Rijswijk;J. Demmers;G. Jenster
目前的肿瘤标志物缺乏敏感性和特异性，因此加强了寻找新的生物标志物的研究工作。人体体液中潜在肿瘤标志物的鉴定受到以下因素的阻碍：对照和患者样本的大变异性和复杂性、费力的生化分析以及所鉴定的蛋白质不太可能由患病细胞产生而是由于次级身体防御机制。在这里提出的一种新方法中，我们通过在前列腺癌异种移植模型中进行蛋白质组学分析来消除这些问题，在该模型中，人前列腺癌细胞在免疫缺陷的裸鼠中形成肿瘤。使用这一概念，根据定义，小鼠血清中存在的可鉴定为人的蛋白质将源自人前列腺癌异种移植物，并且可能具有潜在的诊断和预后价值。使用一维凝胶电泳，液相色谱和质谱，我们确定了肿瘤衍生的人nm 23/核苷二磷酸激酶（NME）在裸鼠的血清中携带雄激素非依赖性人前列腺癌异种移植物PC 339。已知NME涉及几种肿瘤细胞（包括前列腺癌细胞）的转移潜力。此外，我们在荷瘤小鼠的血清中鉴定了六种参与糖酵解的人类酶（果糖-二磷酸醛缩酶A、丙糖-磷酸异构酶、甘油醛-3-磷酸脱氢酶、α烯醇化酶和乳酸脱氢酶A和B）。通过蛋白质印迹法证实携带PC 339的小鼠血清中存在人NME和甘油醛-3-磷酸脱氢酶。虽然这些蛋白质在预测前列腺癌预后方面的推定有用性仍有待确定，但目前的数据表明，我们的方法是一种有前途的工具，可用于集中发现新的前列腺癌生物标志物。
Lack of sensitivity and specificity of current tumor markers has intensified research efforts to find new biomarkers. The identification of potential tumor markers in human body fluids is hampered by large variability and complexity of both control and patient samples, laborious biochemical analyses, and the fact that the identified proteins are unlikely produced by the diseased cells but are due to secondary body defense mechanisms. In a new approach presented here, we eliminate these problems by performing proteomic analysis in a prostate cancer xenograft model in which human prostate cancer cells form a tumor in an immune-incompetent nude mouse. Using this concept, proteins present in mouse serum that can be identified as human will, by definition, originate from the human prostate cancer xenograft and might have potential diagnostic and prognostic value. Using one-dimensional gel electrophoresis, liquid chromatography, and mass spectrometry, we identified tumor-derived human nm23/nucleoside-diphosphate kinase (NME) in the serum of a nude mouse bearing the androgen-independent human prostate cancer xenograft PC339. NME is known to be involved in the metastatic potential of several tumor cells, including prostate cancer cells. Furthermore we identified six human enzymes involved in glycolysis (fructose-bisphosphate aldolase A, triose-phosphate isomerase, glyceraldehyde-3-phosphate dehydrogenase, α enolase, and lactate dehydrogenases A and B) in the serum of the tumor-bearing mice. The presence of human NME and glyceraldehyde-3-phosphate dehydrogenase in the serum of PC339-bearing mice was confirmed by Western blotting. Although the putative usefulness of these proteins in predicting prognosis of prostate cancer remains to be determined, the present data illustrate that our approach is a promising tool for the focused discovery of new prostate cancer biomarkers.