Expression profiling with RNA from formalin-fixed, paraffin-embedded material.
Expression profiling with RNA from formalin-fixed, paraffin-embedded material.
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用福尔马林固定，石蜡包裹的材料的RNA表达分析。
DOI：
10.1186/1755-8794-1-9
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发表时间：
2008-04-19
影响因子：
2.7
通讯作者：
Jaggi R
中科院分区：
文献类型：
--
作者：
Oberli A;Popovici V;Delorenzi M;Baltzer A;Antonov J;Matthey S;Aebi S;Altermatt HJ;Jaggi R
通过基因表达谱对乳腺癌和其他癌症进行分子表征，证实了现有的分类，并揭示了新的亚型。大多数分析研究基于新鲜冷冻（FF）肿瘤材料，其仅可用于有限数量的样本，而数千个肿瘤样本以福尔马林固定、石蜡包埋（FFPE）块的形式存在。不幸的是，源自FFPE材料的RNA被片段化和化学修饰，从而损害通过标准程序的表达测量。从FFPE材料中分离RNA的稳健方案适合于稳定和可重复的基因表达测量（例如通过定量逆转录酶PCR，QPCR）仍然是一个主要挑战。我们提出了一个简单的程序，从FFPE材料的诊断样品的RNA分离。当与优化的cDNA合成方案和对短扩增子具有特异性的TaqMan测定组合使用时，RNA适合于通过QPCR进行表达测量。将FFPE衍生的RNA与从相同肿瘤分离的完整RNA进行比较。根据与ER应答、HER2信号传导和增殖相关的基因计算初步评分。来自FFPE材料的完整和部分片段化RNA之间的相关系数为0.83至0.97。我们开发了一种从FFPE材料中分离RNA的简单而稳健的方法。RNA可用于基因表达谱分析。来自几个基因的表达测量可以组合成代表肿瘤的激素或增殖状态的稳健分数。
Molecular characterization of breast and other cancers by gene expression profiling has corroborated existing classifications and revealed novel subtypes. Most profiling studies are based on fresh frozen (FF) tumor material which is available only for a limited number of samples while thousands of tumor samples exist as formalin-fixed, paraffin-embedded (FFPE) blocks. Unfortunately, RNA derived of FFPE material is fragmented and chemically modified impairing expression measurements by standard procedures. Robust protocols for isolation of RNA from FFPE material suitable for stable and reproducible measurement of gene expression (e.g. by quantitative reverse transcriptase PCR, QPCR) remain a major challenge. We present a simple procedure for RNA isolation from FFPE material of diagnostic samples. The RNA is suitable for expression measurement by QPCR when used in combination with an optimized cDNA synthesis protocol and TaqMan assays specific for short amplicons. The FFPE derived RNA was compared to intact RNA isolated from the same tumors. Preliminary scores were computed from genes related to the ER response, HER2 signaling and proliferation. Correlation coefficients between intact and partially fragmented RNA from FFPE material were 0.83 to 0.97. We developed a simple and robust method for isolating RNA from FFPE material. The RNA can be used for gene expression profiling. Expression measurements from several genes can be combined to robust scores representing the hormonal or the proliferation status of the tumor.
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影响因子：
3.1
作者：
KOOPMANS, M;MONROE, SS;ZAKI, SR
通讯作者：
ZAKI, SR
影响因子：
4.4
作者：
Bibikova, Marina;Chudin, Eugene;Wang-Rodriguez, Jessica
通讯作者：
Wang-Rodriguez, Jessica
影响因子：
6
作者：
Bibikova, M;Talantov, D;Fan, JB
通讯作者：
Fan, JB
DOI：
10.1006/meth.1996.0407
发表时间：
1997-02-01
期刊：
METHODS-A COMPANION TO METHODS IN ENZYMOLOGY
影响因子：
--
作者：
Orlando, V;Strutt, H;Paro, R
通讯作者：
Paro, R
影响因子：
45.3
作者：
Loi, Sherene;Haibe-Kains, Benjamin;Sotiriou, Christos
通讯作者：
Sotiriou, Christos