比较福尔马林固定组织、石蜡包埋组织和qPCR的基因表达数据与速冻组织和微阵列的基因表达数据,以模拟卵巢癌患者的预后。

Q2 Medicine BMC Clinical Pathology Pub Date : 2015-09-24 eCollection Date: 2015-01-01 DOI:10.1186/s12907-015-0017-1
William H Bradley, Kevin Eng, Min Le, A Craig Mackinnon, Christina Kendziorski, Janet S Rader
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引用次数: 13

摘要

背景:之前,我们已经使用来自癌症基因组图谱(TCGA)的临床和基因表达数据来建立一个基于通路的指数来预测卵巢癌的预后。这些数据是通过Affymetrix U133平台测量的速冻组织获得的。在目前的研究中,我们将用于建模的数据与来自TaqMan qPCR的数据联系起来,包括速冻和石蜡包埋(FFPE)样本。方法:为了比较保存方法对qPCR检测基因表达的影响,我们评估了18例患者和肿瘤样本匹配的快速冷冻和FFPE卵巢癌样本。为了比较基因测量技术,我们将10例肿瘤样本匹配速冻卵巢癌样本的qPCR数据与TCGA的微阵列数据进行了关联。我们将结果归一化为三个管家基因的平均表达。我们将数据缩放并居中,以便与Affymetrix输出进行比较。结果:在我们的TaqMan实验中,对于18个标本,从快速冷冻组织中获得的基因表达数据与从FFPE样品中获得的基因表达数据高度相关(r > 0.82)。对于10个重复的TCGA标本,报道的微阵列数据与我们的qPCR数据相关性良好(r = 0.6),并且沿通路的表达范围相似。结论:通过qPCR获得的FFPE浆液性卵巢癌样本基因表达数据可用于基于通路的预测模型的评估。规范化程序描述了表达的控制变化,并且沿着特定途径计算的范围可以解释为患者的风险概况。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

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Comparing gene expression data from formalin-fixed, paraffin embedded tissues and qPCR with that from snap-frozen tissue and microarrays for modeling outcomes of patients with ovarian carcinoma.

Background: Previously, we have used clinical and gene expression data from The Cancer Genome Atlas (TCGA) to model a pathway-based index predicting outcomes in ovarian carcinoma. This data were obtained from snap-frozen tissue measured with the Affymetrix U133 platform. In the current study, we correlate the data used to model with data derived from TaqMan qPCR both snap frozen and paraffin embedded (FFPE) samples.

Methods: To compare the effect of preservation methods on gene expression measured by qPCR, we assessed 18 patient and tumor sample matched snap-frozen and FFPE ovarian carcinoma samples. To compare gene measurement technologies, we correlated qPCR data from 10 patients with tumor sample matched snap-frozen ovarian carcinoma samples with the microarray data from TCGA. We normalized results to the average expression of three housekeeping genes. We scaled and centered the data for comparison to the Affymetrix output.

Results: For the 18 specimens, gene expression data obtained from snap-frozen tissue correlated highly with that from FFPE samples in our TaqMan assay (r > 0.82). For the 10 duplicate TCGA specimens, the reported microarray data correlated well (r = 0.6) with our qPCR data, and ranges of expression along pathways were similar.

Conclusions: Gene expression data obtained by qPCR from FFPE serous ovarian carcinoma samples can be used to assess in the pathway-based predictive model. The normalization procedures described control variations in expression, and the range calculated along a specific pathway can be interpreted for a patient's risk profile.

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来源期刊
BMC Clinical Pathology
BMC Clinical Pathology Medicine-Pathology and Forensic Medicine
CiteScore
3.30
自引率
0.00%
发文量
0
期刊介绍: BMC Clinical Pathology is an open access journal publishing original peer-reviewed research articles in all aspects of histopathology, haematology, clinical biochemistry, and medical microbiology (including virology, parasitology, and infection control). BMC Clinical Pathology (ISSN 1472-6890) is indexed/tracked/covered by PubMed, CAS, EMBASE, Scopus and Google Scholar.
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