Enelund Lars, Nielsen Lise N, Cirera Susanna
Department of Veterinary Clinical and Animal Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen. Denmark.
Microrna. 2017;6(1):42-52. doi: 10.2174/2211536606666170113124114.
Early and specific detection of cancer is of great importance for successful treatment of the disease. New biomarkers, such as microRNAs, could improve treatment efficiency and survival ratio. In human medicine, deregulation of microRNA profiles in circulation has shown great potential as a new type of biomarker for cancer diagnostics. There are, however, few studies of circulating microRNAs in dogs. Extracellular circulating microRNAs have shown a high level of stability in human blood and other body fluids. Nevertheless, there are still important issues to be solved before microRNAs can be applied routinely as a clinical tool, one of them being their stability over time in media commonly used for blood sampling.
Evaluation of the stability of microRNA levels in plasma and serum from healthy dogs after storage at room temperature for different time points before being processed.
The levels of four microRNAs (cfa-let-7a, cfa-miR-16, cfa-miR-23a and cfa-miR-26a) known to be stably expressed from other canine studies, have been measured by quantitative real-time PCR (qPCR).
MicroRNA levels were found sufficiently stable for gene profiling in serum- and plasma stored at room temperature for 1 hour but not for samples stored at room temperature for 24 hours.
Storage at room temperature of serum and plasma samples intended for microRNA profiling should be kept for a minimum period of time before proceeding with RNA isolation. For the four microRNAs investigated in the present study, we did not find significant differences in microRNA levels between serum and plasma samples from the same time point.
癌症的早期特异性检测对于该疾病的成功治疗至关重要。新的生物标志物，如微小RNA，可提高治疗效率和生存率。在人类医学中，循环中微小RNA谱的失调已显示出作为一种新型癌症诊断生物标志物的巨大潜力。然而，关于犬类循环微小RNA的研究很少。细胞外循环微小RNA在人类血液和其他体液中显示出高度稳定性。尽管如此，在微小RNA能够作为临床工具常规应用之前，仍有一些重要问题需要解决，其中之一是它们在常用血液采样介质中的长期稳定性。
评估健康犬的血浆和血清在处理前于室温下储存不同时间点后微小RNA水平的稳定性。
通过定量实时聚合酶链反应（qPCR）测量了已知在其他犬类研究中稳定表达的四种微小RNA（cfa-let-7a、cfa-miR-16、cfa-miR-23a和cfa-miR-26a）的水平。
发现微小RNA水平在室温下储存1小时的血清和血浆中足够稳定，可用于基因谱分析，但在室温下储存24小时的样本中则不然。
用于微小RNA谱分析的血清和血浆样本在室温下储存的时间应尽可能短，然后再进行RNA分离。对于本研究中调查的四种微小RNA，我们未发现同一时间点的血清和血浆样本之间微小RNA水平存在显著差异。
