机器人辅助腹腔镜前列腺切除术和开放性前列腺切除术后用于分子分析的前列腺组织完整性
Integrity of prostatic tissue for molecular analysis after robotic-assisted laparoscopic and open prostatectomy.
作者信息
Best Sara, Sawers Youssef, Fu Vivian X, Almassi Nima, Huang Wei, Jarrard David F
机构信息
Department of Surgery, Division of Urology, University of Wisconsin School of Medicine and Public Health, Madison, Wisconsin 53792, USA.
出版信息
Urology. 2007 Aug;70(2):328-32. doi: 10.1016/j.urology.2007.04.005.
OBJECTIVES
The warm ischemia time of tissue before fixation for pathologic analysis has been linked to changes in cell morphology and nucleic acid and protein integrity. Robotic-assisted laparoscopic prostatectomy (RALP) results in longer warm ischemia times than open radical retropubic prostatectomy. To assess the effect of longer ischemia times on biomolecular integrity, we analyzed DNA, RNA and protein collected from robotic and open prostatectomy specimens.
METHODS
We examined 22 consecutive RALP (n = 11) and open radical retropubic prostatectomy (n = 11) specimens after hematoxylin-eosin staining by light microscopy. To assess protein integrity, immunohistochemical staining for p63, E-cadherin, and AE1/AE3 was performed. DNA was assessed by gel analysis. An RNA integrity score was determined by microfluidic capillary electrophoresis and calculated according to the electropherogram findings and simulated gel view. Finally, epithelial cells were cultured on collagen-coated plates.
RESULTS
No differences in clinicopathologic characteristics were found between the two groups, with the exception of a significantly longer warm ischemia time during RALP (82 +/- 23 minutes) compared with open radical retropubic prostatectomy (23 +/- 2 minutes; P <0.001). Tissue integrity was suitable for the assessment of pathologic grade and stage for all samples. Protein and DNA analyses demonstrated no evidence of degradation in any samples. No significant differences in the RNA integrity scores were demonstrated between the surgical approaches. Prostate epithelial cells were cultured successfully in 66% of the RALP specimens.
CONCLUSIONS
RALP, although it involves additional exposure to warm ischemia, does not significantly affect the histopathologic characteristics or biomolecular integrity of the specimen. Provided a rapid response occurs for tissue banking after specimen removal, molecular research studies using prostatic tissue harvested by way of RALP appear feasible.
目的
用于病理分析的组织在固定前的热缺血时间与细胞形态、核酸及蛋白质完整性的变化有关。机器人辅助腹腔镜前列腺切除术（RALP）导致的热缺血时间比开放性耻骨后根治性前列腺切除术更长。为评估较长缺血时间对生物分子完整性的影响，我们分析了从机器人辅助和开放性前列腺切除标本中收集的DNA、RNA和蛋白质。
方法
我们对22例连续的RALP（n = 11）和开放性耻骨后根治性前列腺切除术（n = 11）标本进行苏木精-伊红染色后，通过光学显微镜检查。为评估蛋白质完整性，进行了p63、E-钙黏蛋白和AE1/AE3的免疫组织化学染色。通过凝胶分析评估DNA。通过微流控毛细管电泳确定RNA完整性评分，并根据电泳图谱结果和模拟凝胶视图进行计算。最后，将上皮细胞培养在胶原包被的培养板上。
结果
两组间临床病理特征无差异，但RALP期间的热缺血时间（82±23分钟）明显长于开放性耻骨后根治性前列腺切除术（23±2分钟；P<0.001）。所有样本的组织完整性均适合评估病理分级和分期。蛋白质和DNA分析表明，任何样本均无降解迹象。两种手术方式之间的RNA完整性评分无显著差异。66%的RALP标本成功培养出前列腺上皮细胞。
结论
RALP虽然涉及额外的热缺血暴露，但对标本的组织病理学特征或生物分子完整性无显著影响。如果在标本切除后对组织库能迅速做出反应，那么使用通过RALP获取的前列腺组织进行分子研究似乎是可行的。
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