Introduction: Personalized treatment, supported by biomarkers, would improve survival of ovarian cancer patients. RNA molecules are potentially important biomarkers. The Danish CancerBiobank provides an infrastructure for handling and storage of biological material, including RNA, from Danish cancer patients. The aim of this study was to investigate the effects of handling-time and fresh-freezing versus RNAlater® fixation on RNA degradation in solid tissue from pelvic mass samples. Materials and Methods: We evaluated RNA quality in surgical tissue from patients with a pelvic mass. Corresponding samples were either fresh-frozen or fixed in RNAlater, at eight different time points after the surgery. Integrity was measured using a bioanalyzer, and the amount and quality were further investigated by quantitative reverse transcription-polymerase chain reaction measuring the expression of housekeeping genes B2M and HPRT1. Results: Our results show that tissue RNA is stable up to at least 180 minutes after the surgery, as the quality was comparable to the quality of RNA handled immediately. Likewise, patient RNA was of acceptable quality after both fresh-frezing and RNAlater fixation, but RNAlater fixation was slightly more effective for RNA preservation. Discussion and Conclusion: Our data suggest that RNA in pelvic mass samples is relatively stable. Knowledge about RNA stability is an important prerequisite for research in RNA biomarkers, where the challenge is to balance the need for careful RNA handling and storage with the need for effective large-scale biobanking in a busy clinical setting where patient treatment is the main priority.

中文翻译：

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优化的用于保存组织中RNA的生物库程序：速冻法和RNAlater固定法的比较。

简介：在生物标志物的支持下进行个性化治疗将提高卵巢癌患者的生存率。RNA分子是潜在的重要生物标志物。丹麦CancerBiobank提供了一个基础设施，用于处理和储存来自丹麦癌症患者的生物材料，包括RNA。这项研究的目的是研究处理时间和新鲜冷冻与RNAlater®固定对盆腔肿块样本中实体组织中RNA降解的影响。材料和方法：我们评估了盆腔肿块患者手术组织中的RNA质量。在手术后的八个不同时间点，将相应的样品新鲜冷冻或固定在RNAlater中。使用生物分析仪测量完整性，通过定量逆转录-聚合酶链反应测定管家基因B2M和HPRT1的表达，进一步研究其数量和质量。结果：我们的结果表明，组织RNA在手术后至少180分钟内是稳定的，因为其质量与立即处理的RNA的质量相当。同样，新鲜冷冻和RNAlater固定后，患者RNA的质量也可以接受，但是RNAlater固定对RNA的保存稍微有效。讨论与结论：我们的数据表明骨盆大量样品中的RNA相对稳定。有关RNA稳定性的知识是进行RNA生物标记物研究的重要先决条件，