Selection of Reference Genes for Normalization of qRT-PCR Data Derived From FFPE Breast Tumors
Selection of Reference Genes for Normalization of qRT-PCR Data Derived From FFPE Breast Tumors
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DOI：
10.1097/pdm.0b013e31817c1ae2
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发表时间：
2009-06-01
影响因子：
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通讯作者：
Dowsett, Mitch
中科院分区：
文献类型：
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作者：
Drury, Suzanne;Anderson, Helen;Dowsett, Mitch
采用实时定量聚合酶链反应对福尔马林固定石蜡包埋（FFPE）材料进行基因表达分析是临床样本回顾性分析的一种有价值的工具。FFPE组织中RNA的降解性质会导致样品间基因表达检测的差异，因此，有必要选择能够反映这种差异的参考基因。为了鉴定FFPE乳腺肿瘤中靶基因标准化的合适参考，在10个内分泌治疗初治的原发性浸润性乳腺肿瘤中测量10个潜在参考的表达。最稳定表达的参考基因与免费提供的算法geNorm的使用进行了鉴定。使用MRPL 19、TBP和TFRC的几何平均值作为FFPE组织的参考因子，FFPE和匹配的冷冻肿瘤之间的ERD α和HER2测量值具有良好的相关性，表明这种参考基因的组合成功地消除了RNA降解对定量实时聚合酶链反应分析的影响。
Gene expression analysis of formalin-fixed paraffin-embedded (FFPE) material by quantitative real-time polymerase chain reaction is a Valuable tool for the retrospective analysis of clinical samples. The degraded nature of RNA from FFPE tissue can lead to variation in detection of gene expression between samples, hence, it is necessary to select reference genes that can reflect this variation. To identify suitable references for the normalization of target genes in FFPE breast tumors, expression of 10 potential references were Measured in 10 endocrine therapy-naive, primary invasive breast tumors. The most stably expressed reference genes were identified with the use of the freely available algorithm geNorm. Using the geometric mean of MRPL19, TBP, and TFRC as a reference factor for FFPE tissue, there was a good correlation of ERD alpha and HER2 measurements between FFPE and matched frozen tumors, indicating that this combination of reference genes successfully removed the effect of RNA degradation on quantitative real-time polymerase chain reaction analysis.