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评估存档的 Bouin 固定石蜡包埋组织标本是否适合蛋白质组学研究。

Evaluation of the suitability of archival Bouin-fixed paraffin-embedded tissue specimens to proteomic investigation.

机构信息

Proteomics Laboratory, Porto Conte Ricerche Srl, Tramariglio, Alghero, Italy.

出版信息

Electrophoresis. 2012 May;33(9-10):1375-84. doi: 10.1002/elps.201200026.

DOI:10.1002/elps.201200026
PMID:22648804
Abstract

Bouin's solution has been used for over a century as a common fixative in several pathology laboratories worldwide. Therefore, a considerable number of Bouin-fixed paraffin-embedded (BFPE) tumor samples of various origin are available in hospital repositories as a powerful information mine for clinical investigations. To date, however, such archived tissues have not been subjected to a systematic study aimed to evaluate their potential use in proteomics. In this report, we investigated whether archival BFPE tissue specimens could be exploited for proteomic studies, upon application of protein extraction and proteomic analysis methods previously optimized for formalin-fixed samples. As a result, gastric BFPE protein extracts exhibited poor suitability for 2D-PAGE analysis, whereas over 300 unique proteins could be successfully detected when extracts were subjected to SDS-PAGE followed by LC-MS/MS (GeLC-MS/MS). Among these, several known markers for gastric cancer and normal gastric functionality were identified, indicative of biological and clinical significance of proteomic data mined from BFPE tissues. A quantitative and qualitative comparison of FFPE and BFPE tissue proteomes was also performed, and results are reported. In conclusion, we demonstrated that BFPE specimens can be analyzed by means of a proteomic approach such as GeLC-MS/MS. Although considerable molecular biases and technical constraints exist, BFPE tissue archives can be fruitfully exploited for gathering proteomic data from particularly precious samples.

摘要

布恩氏溶液作为一种常用的固定剂,在全球范围内的多个病理学实验室中使用了一个多世纪。因此,医院的储存库中保存了大量来自不同来源的布恩氏固定石蜡包埋(BFPE)肿瘤样本,这些样本是临床研究的宝贵信息来源。然而,到目前为止,这些存档组织尚未经过系统研究,以评估它们在蛋白质组学中的潜在用途。在本报告中,我们研究了在应用先前针对福尔马林固定样本优化的蛋白质提取和蛋白质组分析方法的情况下,存档的 BFPE 组织标本是否可用于蛋白质组学研究。结果表明,胃 BFPE 蛋白质提取物不适于 2D-PAGE 分析,而当提取物经过 SDS-PAGE 后进行 LC-MS/MS(GeLC-MS/MS)分析时,可以成功检测到 300 多种独特的蛋白质。其中,鉴定出了几种用于胃癌和正常胃功能的已知标志物,表明从 BFPE 组织中挖掘的蛋白质组学数据具有生物学和临床意义。还对 FFPE 和 BFPE 组织蛋白质组进行了定量和定性比较,并报告了结果。总之,我们证明了 BFPE 标本可以通过 GeLC-MS/MS 等蛋白质组学方法进行分析。尽管存在相当大的分子偏见和技术限制,但 BFPE 组织档案可以成功地用于从特别珍贵的样本中收集蛋白质组学数据。

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