Hammer Elke, Ernst Florian D, Thiele Andrea, Karanam Narasimha Kumar, Kujath Christina, Evert Matthias, Völker Uwe, Barthlen Winfried
Interfaculty Institute for Genetics and Functional Genomics, Department of Functional Genomics, University Medicine Greifswald, Friedrich-Ludwig-Jahn-Str. 15a, D-17475 Greifswald, Germany.
Interfaculty Institute for Genetics and Functional Genomics, Department of Functional Genomics, University Medicine Greifswald, Friedrich-Ludwig-Jahn-Str. 15a, D-17475 Greifswald, Germany; Clinic for Pediatric Surgery, University Medicine Greifswald, Sauerbruchstr. 1, D-17475 Greifswald, Germany.
Clin Chim Acta. 2014 Jun 10;433:235-41. doi: 10.1016/j.cca.2014.03.020. Epub 2014 Mar 27.
Wilms' tumor (nephroblastoma, WT) is the most frequent renal cancer in children. However, molecular details leading to WT have not been characterized sufficiently yet. Proteomic studies might provide new insights but are hampered by limited availability of fresh frozen tissue specimen. Therefore, we tested formalin-fixed paraffin-embedded (FFPE) tissue sections routinely collected for pathological inspection for their use in in-depth-proteomic analyses of WT samples in comparison to fresh frozen specimen. The overlap of the proteins identified was over 65%. Thus we used FFPE material from 7 patients for tandem mass spectrometry based comparison of the proteomes of WT and healthy renal tissues. We detected 262 proteins, which were differentially expressed in tumor compared to healthy renal tissue. The majority of these proteins displayed lower levels in the tumor tissue and only 30% higher levels. For selected candidates data were confirmed by immunohistochemical staining. Correlation analysis of blastemal proportions in WT and protein intensities revealed candidates for tumor stratification.
This proof of principle proteomic study of FFPE tissue sections from WT patients demonstrates that these archived tissues constitute a valuable resource for larger in-depth proteomic studies to identify markers to follow chemotherapy efficiency or for stratification of tumor subtypes.
肾母细胞瘤(WT)是儿童中最常见的肾癌。然而,导致WT的分子细节尚未得到充分表征。蛋白质组学研究可能会提供新的见解,但由于新鲜冷冻组织样本的可用性有限而受到阻碍。因此,我们测试了常规收集用于病理检查的福尔马林固定石蜡包埋(FFPE)组织切片,以用于WT样本的深入蛋白质组学分析,并与新鲜冷冻样本进行比较。鉴定出的蛋白质重叠率超过65%。因此,我们使用了7例患者的FFPE材料,通过串联质谱对WT和健康肾组织的蛋白质组进行比较。我们检测到262种蛋白质,它们在肿瘤组织中与健康肾组织相比有差异表达。这些蛋白质中的大多数在肿瘤组织中水平较低,只有30%的蛋白质水平较高。对于选定的候选蛋白,通过免疫组织化学染色对数据进行了确认。WT中胚层比例与蛋白质强度的相关性分析揭示了肿瘤分层的候选蛋白。
这项对WT患者FFPE组织切片进行的原理验证蛋白质组学研究表明,这些存档组织是进行更大规模深入蛋白质组学研究的宝贵资源,可用于识别跟踪化疗疗效的标志物或肿瘤亚型的分层。