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BMC Clin Pathol. 2015 Sep 24;15:17. doi: 10.1186/s12907-015-0017-1. eCollection 2015.

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.

BMC clinical pathology

William H Bradley, Kevin Eng, Min Le, A Craig Mackinnon, Christina Kendziorski, Janet S Rader

Affiliations

  1. Department of Obstetrics and Gynecology, Medical College of Wisconsin, 8701 Watertown Plank Road, Milwaukee, WI 53226 USA.
  2. Department of Biostatistics and Medical Informatics, University of Wisconsin-Madison, Madison, WI 53792 USA ; Current Address: Department of Biostatistics and Bioinformatics, Roswell Park Cancer Institute, Buffalo, NY USA.
  3. Department of Pathology, Medical College of Wisconsin, Milwaukee, WI 53226 USA.
  4. Department of Biostatistics and Medical Informatics, University of Wisconsin-Madison, Madison, WI 53792 USA.

PMID: 26412982 PMCID: PMC4582729 DOI: 10.1186/s12907-015-0017-1

Abstract

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.

References

  1. PLoS One. 2011 Feb 11;6(2):e17163 - PubMed
  2. Histochem Cell Biol. 2008 Sep;130(3):435-45 - PubMed
  3. Gynecol Oncol. 2013 Apr;129(1):159-64 - PubMed
  4. Nat Biotechnol. 2006 Sep;24(9):1115-22 - PubMed
  5. Stat Med. 2013 Apr 30;32(9):1524-35 - PubMed
  6. Nat Biotechnol. 2006 Sep;24(9):1151-61 - PubMed
  7. BMC Bioinformatics. 2008 Aug 12;9 Suppl 9:S10 - PubMed
  8. Nat Rev Drug Discov. 2008 Nov;7(11):875-6 - PubMed
  9. N Engl J Med. 2012 Mar 8;366(10):883-92 - PubMed
  10. Nature. 2011 Jun 29;474(7353):609-15 - PubMed
  11. Science. 2008 Sep 26;321(5897):1801-6 - PubMed
  12. BMC Genomics. 2008 Jul 11;9:328 - PubMed
  13. BMC Med Genomics. 2009 May 08;2:23 - PubMed
  14. BMC Genomics. 2006 Mar 21;7:59 - PubMed
  15. Pathol Res Pract. 2013 Dec;209(12):784-9 - PubMed
  16. Nat Methods. 2005 May;2(5):345-50 - PubMed

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