Proteomic analysis of formalin-fixed prostate cancer tissue
Proteomic analysis of formalin-fixed prostate cancer tissue
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DOI：
10.1074/mcp.m500102-mcp200
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发表时间：
2005-11-01
影响因子：
7
通讯作者：
Krizman, DB
中科院分区：
文献类型：
--
作者：
Hood, BL;Darfler, MM;Krizman, DB
福尔马林固定石蜡包裹（FFPE）组织的蛋白质组学分析将使对全球存在的庞大的病理表征临床样品的庞大档案进行回顾性生物标志物研究。然而，这些FFPE组织使用许多艺术方法的蛋白质组学研究是难治性的，这在很大程度上是由于福尔马林固定引起的高量共价交联蛋白。已经开发了一种新型的组织显微解剖技术，并与一种直接从FFPE组织中提取可溶性肽的方法结合使用，以进行前列腺癌（PCA）和良性前列腺增生（BPH）的质谱分析。鉴定了来自PCA和BPH组织的数百种蛋白质，包括几种已知的PCA标记，例如前列腺特异性抗原，前列腺酸性磷酸酶和巨噬细胞抑制性细胞因子1。利用稳定的同位素标记的定量蛋白质组学分析证实，在BPH和PCA细胞中，前列腺特异性抗原和前列腺酸磷酸酶的表达水平相似，而与BPH细胞相比，PCA中发现巨噬细胞抑制性细胞因子-1的表达更大。
Proteomic analysis of formalin-fixed paraffin-embedded (FFPE) tissue would enable retrospective biomarker investigations of this vast archive of pathologically characterized clinical samples that exist worldwide. These FFPE tissues are, however, refractory to proteomic investigations utilizing many state of the art methodologies largely due to the high level of covalently cross-linked proteins arising from formalin fixation. A novel tissue microdissection technique has been developed and combined with a method to extract soluble peptides directly from FFPE tissue for mass spectral analysis of prostate cancer (PCa) and benign prostate hyperplasia (BPH). Hundreds of proteins from PCa and BPH tissue were identified, including several known PCa markers such as prostate-specific antigen, prostatic acid phosphatase, and macrophage inhibitory cytokine-1. Quantitative proteomic profiling utilizing stable isotope labeling confirmed similar expression levels of prostate-specific antigen and prostatic acid phosphatase in BPH and PCa cells, whereas the expression of macrophage inhibitory cytokine-1 was found to be greater in PCa as compared with BPH cells.