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. 2014 Jan;74(1):61-9.
doi: 10.1002/pros.22730. Epub 2013 Sep 21.

Biobanking of derivatives from radical retropubic and robot-assisted laparoscopic prostatectomy tissues as part of the prostate cancer biorepository network

Medha Darshan  1 , Qizhi Zheng, Helen L Fedor, Nicolas Wyhs, Srinivasan Yegnasubramanian, Peng Lee, Jonathan Melamed, George J Netto, Bruce J Trock, Angelo M De Marzo, Karen S Sfanos
Affiliations

Biobanking of derivatives from radical retropubic and robot-assisted laparoscopic prostatectomy tissues as part of the prostate cancer biorepository network

Medha Darshan et al. Prostate. 2014 Jan.

Abstract

Background: The goal of the Prostate Cancer Biorepository Network (PCBN) is to develop a biorepository with high-quality, well-annotated specimens obtained in a systematic, reproducible fashion using optimized and standardized protocols, and an infrastructure to facilitate the growth of the resource and its wide usage by the prostate cancer research community. An emerging area of concern in the field of prostate cancer biobanking is an apparent shift in the proportion of surgical procedures performed for prostate cancer treatment from radical retropubic prostatectomy (RRP) to robot-assisted laparoscopic prostatectomy (RALP). Our study aimed to determine the potential impact of the RALP procedure on the detection of known prostate cancer biomarkers, and the subsequent suitability of RALP-derived specimens for prostate cancer biomarker studies.

Methods: DNA and RNA were extracted from RRP and RALP specimens. Quality assessment was conducted using spectrophotometric analysis and RNA was analyzed for RNA integrity number (RIN) and by real-time reverse-transcription PCR (qRT-PCR) for racemase, hepsin, ERG, TMPRSS2-ERG gene fusions, and the microRNAs miR-26a, miR-26b, miR-141, and miR-221.

Results: We demonstrate that extraction of derivatives from frozen tissues from RRP and RALP specimens yields samples of equally high quality as assessed by spectrophotometric and RIN analysis. Likewise, expression levels of genes analyzed by qRT-PCR did not differ between RRP and RALP-derived tissues.

Conclusions: Our studies indicate that samples obtained from RALP specimens may be suitable for prostate cancer biomarker studies-an important finding given the current shift in surgical procedures for prostate cancer treatment.

Keywords: DNA; RNA; biobanking; biomarker; biorepository; prostate cancer.

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Figures

Fig. 1
Fig. 1
Assessment of nucleic acid quality as part of SOP development. A: Agarose gel of DNA samples extracted from frozen prostate tissue specimens. B: RIN of RNA samples extracted from frozen prostate tissues (Agilent Bioanalyzer). Mean RIN = 7.7 ± 1.8, median RIN = 8.3.
Fig. 2
Fig. 2
Distribution of RIN numbers obtained for RNA extracted from radical retropubic prostatectomy (RRP, n = 83) versus robot-assisted laparoscopic prostatectomy (RALP, n = 20) specimens. Median RIN for RRP versus RALP specimens was 8.3 (interquartile range 7.3–8.8) and 8.5 (interquartile range 7.7–8.6), respectively, P = 0.923 (Wilcoxon rank sum test).
Fig. 3
Fig. 3
Fold difference in racemase and hepsin expression in tumor as compared to benign among tumor/benign RNA pairs from RRP and RALP.
Fig. 4
Fig. 4
Fold difference in ERG expression in tumor samples as compared to benign among RNA pairs from RRP and RALP. Samples shown in grey represent cases where the tumor sample was found to be positive for TMPRSS2-ERG gene fusions via qPCR.
Fig. 5
Fig. 5
Fold difference in miR-26a, miR-26b, miR-141, and miR-221 expression in tumor samples relative to benign among RNA pairs from RRP and RALP as assessed by TaqMan® small RNA assay.
See this image and copyright information in PMC

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