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Assessing the Impact of Tissue Devitalization Time on Genome-wide Gene Expression Analysis in Ovarian Tumor Samples

Dumur, Catherine I. PhD*; Sana, Sherjeel MD; Ladd, Amy C. PhD*; Ferreira-Gonzalez, Andrea PhD*; Wilkinson, David S. MD, PhD*; Powers, Celeste N. MD, PhD*; Garrett, Carleton T. MD, PhD*

Diagnostic Molecular Pathology: December 2008 - Volume 17 - Issue 4 - pp 200-206
doi: 10.1097/PDM.0b013e318169bfaf
Original Articles
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The utilization of genome-wide gene expression microarray technology in tumor stratification has proven a powerful tool to identify gene expression signatures associated with cancer prognosis and is currently under evaluation in clinical laboratories. Standardized protocols, including tumor tissue postoperatively handling guidelines are yet to be defined. We aimed at assessing a systematic effect of devitalization in ovarian tumors' gene expression profiling, using high-density oligonucleotide microarrays, under a standardized protocol following strict quality control criteria. Residual tissue from the surgical pathology specimen was divided into 5 samples. Half of each was immediately snap frozen in liquid nitrogen. The remaining halves were kept at room temperature for 0, 15, 30, 60, and 120 minutes, at which time the tissue was snap frozen in liquid nitrogen, and stored at −80°C until RNA extraction. The entire process from RNA extraction through feature intensity distribution was rigorously monitored for quality. Identification of altered gene expression among each pair of snap frozen and devitalized samples per ovarian tumor specimen was assessed by using the Significance score (S-score) method. We identified only 4 probe sets that seemed to correlate with devitalization time in one of the ovarian tumor specimens, suggesting that they are not likely to have an impact on gene expression profiling tumor stratification. Our study suggests that with proper sample handling and rigorous quality control procedures for RNA extraction and microarray analysis, tumor classification based on global gene expression data will not be adversely affected if devitalization times are kept within a 120-minute window.

*Department of Pathology, Virginia Commonwealth University, Richmond, VA

Department of Internal Medicine, Southern Illinois University School of Medicine, Springfield, IL

Supported by the Commonwealth Technology Research Fund (CTRF ♯SE2002_02) and the Virginia Tobacco Settlement Foundation.

Reprints: Catherine I. Dumur, PhD, Department of Pathology, Virginia Commonwealth University, Clinical Support Center, Room 247, 403 North 13th Street, Richmond, VA 23298 (e-mail: cdumur@mcvh-vcu.edu).

© 2008 Lippincott Williams & Wilkins, Inc.