FFPE tissue as a feasible source for gene expression analysis - A comparison of three reference genes and one tumor marker
FFPE tissue as a feasible source for gene expression analysis - A comparison of three reference genes and one tumor marker
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DOI：
10.1016/j.prp.2013.09.007
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发表时间：
2013-01-01
影响因子：
2.8
通讯作者：
Hager, Thomas
中科院分区：
文献类型：
--
作者：
Walter, Robert Fred Henry;Mairinger, Fabian Dominik;Hager, Thomas
背景：福尔马林固定、石蜡包埋是现代病理学中肿瘤组织的标准处理技术。低温保存等新技术允许快速固定和长期存储，但随之而来的是成本增加和存储复杂性增加。然而，福尔马林固定的石蜡包埋(FFPE)组织大量可用，使其成为回顾研究的理想材料。材料和方法：收集2005-2012年埃森大学医院病理与神经病理研究所收集的80例肺肿瘤标本，采用TaqMan(R)基因按需表达分析方法对其基因表达进行分析。对三个不同的潜在参考基因(ACTB、GAPDH、HPRT1)的表达进行了评估，并将蛋白酶体亚单位(PSMA1)作为肿瘤标志物并作为内部技术对照。结论：对于GAPDH和ACTB，被调查的实体之间存在高度显著的相关性和一致的表达，为进一步研究提供了可靠的参考基因。此外，还验证了基于FFPE组织的基因表达分析的可行性，表明所获得的mRNA质量是足够的。如果仔细进行标准化的FFPE制备，就可以分离出足够的mRNA，用于可靠和成功的基因表达分析。这为大型回溯性研究提供了基础，这些研究将分子和临床随访数据联系起来。(C)2013年爱思唯尔股份有限公司。版权所有。
Background: Formalin-fixation, paraffin-embedding is the standard processing technique for tumor tissue in modern pathology. New techniques such as cryo-conservation allow rapid fixation and long-time storage but come along with increased costs and enlarged storage complexity. However, formalin-fixed, paraffin-embedded (FFPE) tissue is available in a large quantity, making it the ideal material for retrospective studies.The following study was designed to investigate the influence of formalin-fixation on the quality of mRNA and applicability of FFPE-derived mRNA for gene expression analysis. Three potential reference genes for pulmonary tumors with neuroendocrine differentiation were included and tested for their robust expression.Materials and methods: Eighty specimens collected from 2005 to 2012 at the Institute of Pathology and Neuropathology at the University Hospital Essen were analyzed for their gene expression by using TaqMan (R) gene expression assays on demand (AoD). Three distinct potential reference genes (ACTB, GAPDH, HPRT1) were evaluated for their expression, and a proteasome subunit (PSMA1) was included in the analysis as tumor marker and functioned as an internal technical control.Conclusion: For GAPDH and ACTB, a highly significant correlation and consistent expression between the investigated entities was found, making them reliable reference genes for further research. Additionally, the feasibility for a FFPE tissue-based gene expression analysis was verified by showing that the mRNA quality is sufficient. When standardized FFPE preparation is performed carefully, sufficient mRNA can be isolated for reliable and successful gene expression analysis. That provides the basis the door for large, retrospective studies that correlate molecular and clinical follow-up data. (C) 2013 Elsevier GmbH. All rights reserved.