Cargando…

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 cor...

Descripción completa

Detalles Bibliográficos
Autores principales: Bradley, William H., Eng, Kevin, Le, Min, Mackinnon, A. Craig, Kendziorski, Christina, Rader, Janet S.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4582729/
https://www.ncbi.nlm.nih.gov/pubmed/26412982
http://dx.doi.org/10.1186/s12907-015-0017-1
_version_ 1782391746889515008
author Bradley, William H.
Eng, Kevin
Le, Min
Mackinnon, A. Craig
Kendziorski, Christina
Rader, Janet S.
author_facet Bradley, William H.
Eng, Kevin
Le, Min
Mackinnon, A. Craig
Kendziorski, Christina
Rader, Janet S.
author_sort Bradley, William H.
collection PubMed
description 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. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12907-015-0017-1) contains supplementary material, which is available to authorized users.
format Online
Article
Text
id pubmed-4582729
institution National Center for Biotechnology Information
language English
publishDate 2015
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-45827292015-09-26 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 Bradley, William H. Eng, Kevin Le, Min Mackinnon, A. Craig Kendziorski, Christina Rader, Janet S. BMC Clin Pathol Research Article 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. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12907-015-0017-1) contains supplementary material, which is available to authorized users. BioMed Central 2015-09-24 /pmc/articles/PMC4582729/ /pubmed/26412982 http://dx.doi.org/10.1186/s12907-015-0017-1 Text en © Bradley et al. 2015 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research Article
Bradley, William H.
Eng, Kevin
Le, Min
Mackinnon, A. Craig
Kendziorski, Christina
Rader, Janet S.
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
title 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
title_full 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
title_fullStr 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
title_full_unstemmed 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
title_short 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
title_sort 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
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4582729/
https://www.ncbi.nlm.nih.gov/pubmed/26412982
http://dx.doi.org/10.1186/s12907-015-0017-1
work_keys_str_mv AT bradleywilliamh comparinggeneexpressiondatafromformalinfixedparaffinembeddedtissuesandqpcrwiththatfromsnapfrozentissueandmicroarraysformodelingoutcomesofpatientswithovariancarcinoma
AT engkevin comparinggeneexpressiondatafromformalinfixedparaffinembeddedtissuesandqpcrwiththatfromsnapfrozentissueandmicroarraysformodelingoutcomesofpatientswithovariancarcinoma
AT lemin comparinggeneexpressiondatafromformalinfixedparaffinembeddedtissuesandqpcrwiththatfromsnapfrozentissueandmicroarraysformodelingoutcomesofpatientswithovariancarcinoma
AT mackinnonacraig comparinggeneexpressiondatafromformalinfixedparaffinembeddedtissuesandqpcrwiththatfromsnapfrozentissueandmicroarraysformodelingoutcomesofpatientswithovariancarcinoma
AT kendziorskichristina comparinggeneexpressiondatafromformalinfixedparaffinembeddedtissuesandqpcrwiththatfromsnapfrozentissueandmicroarraysformodelingoutcomesofpatientswithovariancarcinoma
AT raderjanets comparinggeneexpressiondatafromformalinfixedparaffinembeddedtissuesandqpcrwiththatfromsnapfrozentissueandmicroarraysformodelingoutcomesofpatientswithovariancarcinoma