Davidsson Sabina, Jerlström Tomas, Carlsson Jessica
Department of Urology, Faculty of Medicine and Health, Örebro University, 703 62 Örebro, Sweden.
Int J Mol Sci. 2025 Sep 3;26(17):8581. doi: 10.3390/ijms26178581.
The reliability of molecular diagnostic and prognostic tools is contingent on the quality of biospecimens, which are often collected during surgical procedures. This study investigated the impact of surgical manipulation on gene expression in the urinary bladder mucosa during radical cystectomy. Seventeen patients with urinary bladder cancer were enrolled, and paired pre- and post-surgery biopsies were analyzed. Pre-surgical biopsies were obtained in situ under anesthesia, while post-surgical biopsies were collected ex vivo following bladder removal. Total RNA was extracted, and gene expression was assessed using qPCR arrays, measuring the expression of 374 inflammation-related genes. The findings from the exploratory phase were further validated by analyzing key genes in an independent patient cohort using TaqMan gene-specific assays. Exploratory analysis revealed significant differential expression in 27 genes, with key genes such as IL6, FOS, and PTGS2 being upregulated post-surgery. Validation of five selected genes in an independent cohort confirmed these findings. This study reinforces the necessity of accounting for surgery-induced alterations in gene expression when analyzing tissue samples collected intraoperatively. By elucidating the molecular impact of surgical interventions, this work provides critical insights for refining experimental methodologies and enhancing the interpretability of gene expression studies in clinical and research settings.
分子诊断和预后工具的可靠性取决于生物样本的质量，这些样本通常在手术过程中采集。本研究调查了根治性膀胱切除术中手术操作对膀胱黏膜基因表达的影响。招募了17例膀胱癌患者，并对手术前后的配对活检样本进行分析。术前活检样本在麻醉下原位获取，术后活检样本在膀胱切除后离体采集。提取总RNA，并使用qPCR阵列评估基因表达，检测374个炎症相关基因的表达。通过使用TaqMan基因特异性检测法分析独立患者队列中的关键基因，对探索阶段的结果进行了进一步验证。探索性分析显示27个基因存在显著差异表达，术后关键基因如IL6、FOS和PTGS2上调。在独立队列中对五个选定基因的验证证实了这些结果。本研究强调，在分析术中采集的组织样本时，必须考虑手术引起的基因表达变化。通过阐明手术干预的分子影响，这项工作为完善实验方法和提高临床及研究环境中基因表达研究的可解释性提供了关键见解。
