Abstract
BackgroundRNA quality is believed to decrease with ischaemia time, and therefore open radical prostatectomy has been advantageous in allowing the retrieval of the prostate immediately after its devascularization. In contrast, robotic-assisted laparoscopic radical prostatectomies (RALP) require the completion of several operative steps before the devascularized prostate can be extirpated, casting doubt on the validity of this technique as a source for obtaining prostatic tissue. We seek to establish the integrity of our biobanking process by measuring the RNA quality of specimens derived from robotic-assisted laparoscopic radical prostatectomy.MethodsWe describe our biobanking process and report the RNA quality of prostate specimens using advanced electrophoretic techniques (RNA Integrity Numbers, RIN). Using multivariate regression analysis we consider the impact of various clinicopathological correlates on RNA integrity.ResultsOur biobanking process has been used to acquire 1709 prostates, and allows us to retain approximately 40% of the prostate specimen, without compromising the histopathological evaluation of patients. We collected 186 samples from 142 biobanked prostates, and demonstrated a mean RIN of 7.25 (standard deviation 1.64) in 139 non-stromal samples, 73% of which had a RIN ≥ 7. Multivariate regression analysis revealed cell type - stromal/epithelial and benign/malignant - and prostate volume to be significant predictors of RIN, with unstandardized coefficients of 0.867(p = 0.001), 1.738(p < 0.001) and -0.690(p = 0.009) respectively. A mean warm ischaemia time of 120 min (standard deviation 30 min) was recorded, but multivariate regression analysis did not demonstrate a relationship with RIN within the timeframe of the RALP procedure.ConclusionsWe demonstrate the robustness of our protocol - representing the concerted efforts of dedicated urology and pathology departments - in generating RNA of sufficient concentration and quality, without compromising the histopathological evaluation and diagnosis of patients. The ischaemia time associated with our prostatectomy technique using a robotic platform does not negatively impact on biobanking for RNA studies.
Highlights
Prostate cancer remains the most common non-dermatological malignancy in men in the Western world [1]
We report the methodology of tissue collection in our radical prostatectomies (RALP) prostate cancer biobank involving 1709 radical prostatectomy specimens
In the rare event that an area of suspicion is only identified at the border of a prostate section, the adjacent biobanked section can be retrieved from storage for further study by the pathologist. 9% of our patients have more tissue taken from the biobank after identifying suspicious areas on clinical specimens, and we believe that the ability to access the biobanked tissue is fundamental to ensuring the integrity of the histological diagnosis
Summary
Prostate cancer remains the most common non-dermatological malignancy in men in the Western world [1]. In order to draw meaningful conclusions from these transcriptomes, investigators must possess robust methods for tissue biobanking, as well as regularly perform quality control on the samples they collect. Variations in both biobanking protocols and quality control methods can limit comparisons between different research groups, and the veracity of any conclusions drawn from their molecular profiles. Robotic-assisted laparoscopic radical prostatectomies (RALP) require the completion of several operative steps before the devascularized prostate can be extirpated, casting doubt on the validity of this technique as a source for obtaining prostatic tissue. We seek to establish the integrity of our biobanking process by measuring the RNA quality of specimens derived from robotic-assisted laparoscopic radical prostatectomy
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