Department of Urology, The First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China.
Asian J Androl. 2012 Sep;14(5):708-14. doi: 10.1038/aja.2012.54. Epub 2012 Aug 6.
Current surgical treatment of prostate cancer is typically accomplished by either open radical prostatectomy (ORP) or robotic-assisted laparoscopic radical prostatectomy (RALRP). Intra-operative procedural differences between the two surgical approaches may alter the molecular composition of resected surgical specimens, which are indispensable for molecular analysis and biomarker evaluation. The objective of this study is to investigate the effect of different surgical procedures on RNA quality and genome-wide expression signature. RNA integrity number (RIN) values were compared between total RNA samples extracted from consecutive LRP (n=11) and ORP (n=24) prostate specimens. Expression profiling was performed using the Agilent human whole-genome expression microarrays. Expression differences by surgical type were analyzed by Volcano plot analysis and gene ontology analysis. Quantitative reverse transcription (RT)-PCR was used for expression validation in an independent set of LRP (n=8) and ORP (n=8) samples. The LRP procedure did not compromise RNA integrity. Differential gene expression by surgery types was limited to a small subset of genes, the number of which was smaller than that expected by chance. Unexpectedly, this small subset of differentially expressed genes was enriched for those encoding transcription factors, oxygen transporters and other previously reported surgery-induced stress-response genes, and demonstrated unidirectional reduction in LRP specimens in comparison to ORP specimens. The effect of the LRP procedure on RNA quality and genome-wide transcript levels is negligible, supporting the suitability of LRP surgical specimens for routine molecular analysis. Blunted in vivo stress response in LRP specimens, likely mediated by CO(2) insufflation but not by longer ischemia time, is manifested in the reduced expression of stress-response genes in these specimens.
目前，前列腺癌的外科治疗通常通过开放性根治性前列腺切除术（ORP）或机器人辅助腹腔镜根治性前列腺切除术（RALRP）来完成。两种手术方法的术中操作差异可能会改变切除手术标本的分子组成，这对于分子分析和生物标志物评估是必不可少的。本研究旨在探讨不同手术程序对 RNA 质量和全基因组表达特征的影响。对连续的 LRP（n=11）和 ORP（n=24）前列腺标本中提取的总 RNA 样本的 RNA 完整性数（RIN）值进行了比较。使用 Agilent 人类全基因组表达微阵列进行表达谱分析。通过火山图分析和基因本体分析对手术类型的表达差异进行了分析。在独立的 LRP（n=8）和 ORP（n=8）样本中使用定量逆转录（RT）-PCR 进行表达验证。LRP 手术不会损害 RNA 完整性。手术类型的差异表达基因仅限于一小部分基因，其数量小于预期的随机数量。出乎意料的是，这些差异表达基因的一小部分富集了那些编码转录因子、氧气转运蛋白和其他先前报道的手术诱导应激反应基因的基因，并且与 ORP 标本相比，LRP 标本中这些基因的表达呈单向减少。LRP 手术对 RNA 质量和全基因组转录水平的影响可以忽略不计，支持 LRP 手术标本适合常规分子分析。LRP 标本中 CO2 充气介导但不是更长的缺血时间介导的体内应激反应减弱，表现在这些标本中应激反应基因的表达减少。
