Lefrak Center of Robotic Surgery & Institute for Prostate Cancer, Brady Foundation Department of Urology, Weill Cornell Medical College, New York, NY, USA.
J Transl Med. 2011 Jul 26;9:121. doi: 10.1186/1479-5876-9-121.
RNA 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.
We 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.
Our 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.
We 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.
人们认为 RNA 的质量会随着缺血时间的延长而降低，因此开放性根治性前列腺切除术有利于在前列腺血管化后立即切除前列腺。相比之下，机器人辅助腹腔镜根治性前列腺切除术（RALP）在可以切除去血管化的前列腺之前需要完成几个手术步骤，这使得该技术作为获取前列腺组织的来源的有效性受到质疑。我们试图通过测量源自机器人辅助腹腔镜根治性前列腺切除术的标本的 RNA 质量来确定我们的生物银行流程的完整性。
我们描述了我们的生物银行流程，并使用先进的电泳技术（RNA 完整性编号，RIN）报告前列腺标本的 RNA 质量。我们使用多元回归分析考虑了各种临床病理相关性对 RNA 完整性的影响。
我们的生物银行流程已用于获取 1709 个前列腺，并且能够保留大约 40%的前列腺标本，而不会影响患者的组织病理学评估。我们从 142 个生物银行前列腺中收集了 186 个样本，在 139 个非基质样本中平均 RIN 为 7.25（标准差 1.64），其中 73%的 RIN ≥ 7。多元回归分析显示细胞类型 - 基质/上皮和良性/恶性 - 和前列腺体积是 RIN 的显著预测因子，未标准化系数分别为 0.867（p = 0.001）、1.738（p < 0.001）和 -0.690（p = 0.009）。记录的平均热缺血时间为 120 分钟（标准差 30 分钟），但多元回归分析在 RALP 手术时间范围内未显示与 RIN 之间存在关系。
我们证明了我们的方案的稳健性 - 代表了专注的泌尿科和病理学部门的共同努力 - 在不影响患者的组织病理学评估和诊断的情况下，产生具有足够浓度和质量的 RNA。与我们使用机器人平台进行的前列腺切除术技术相关的缺血时间不会对 RNA 研究的生物银行产生负面影响。
