Kupec Tomas, Bleilevens Andreas, Iborra Séverine, Najjari Laila, Wittenborn Julia, Maurer Jochen, Stickeler Elmar
Clinic for Gynaecology and Obstetrics, University Hospital RWTH Aachen, Aachen, Germany.
PLoS One. 2022 Aug 31;17(8):e0268958. doi: 10.1371/journal.pone.0268958. eCollection 2022.
There is a strong body of evidence by several translational studies which demonstrate the potential of circulating miRNAs as a potential biomarker in oncology. However, recent reports documented varying stability of these small RNA molecules in serum samples. The aim of our pilot study was to evaluate the stability of miRNAs in serum in relation to food intake and sample storage. Serum miRNA expression levels of 16 different miRNAs from 8 healthy volunteers were quantified by real-time PCR. 4 samples from each donor were analysed-2 samples (fasting, in the morning and after food intake, at noon) were analysed within 24h and 2 samples (fasting and after food intake, at noon) were stored at -80°C for 14 days and subsequently analysed. Student´s t-test was used to determine significant differences. The detectability of the distinct miRNA as a surrogate for the stability of these small RNA molecules was slightly altered by the storage conditions, but only a miRNA 22-3p, out of the analysed 16 miRNAs, shows significant lower dCq expression (3.821 vs. 4.530; p<0,01) by qPCR dependent on storage conditions (-80°C vs. 4°C). However, miRNA levels were not affected by food intake. The difference between samples taken in the morning (fasting) and at noon (after a normal meal) did not show any significant differences. MiRNAs can be considered to be a relatively stable tool in laboratory diagnostics, but clearly every new assay needs thorough evaluation. The stability of miRNAs documented here in healthy volunteers shows their potential in the search for innovative biomarkers in oncology.
多项转化研究有大量证据表明，循环微小RNA（miRNA）有潜力成为肿瘤学中的生物标志物。然而，最近的报告记录了这些小RNA分子在血清样本中的稳定性存在差异。我们的初步研究旨在评估血清中miRNA相对于食物摄入和样本储存的稳定性。通过实时聚合酶链反应（PCR）对8名健康志愿者的16种不同miRNA的血清表达水平进行了定量分析。对每个捐赠者的4个样本进行了分析——2个样本（早晨空腹和中午进食后）在24小时内进行分析，另外2个样本（空腹和中午进食后）在-80°C下储存14天，随后进行分析。采用学生t检验来确定显著差异。作为这些小RNA分子稳定性替代指标的不同miRNA的可检测性受储存条件的影响略有改变，但在分析的16种miRNA中，只有miRNA 22 - 3p在依赖于储存条件（-80°C与4°C）的定量PCR中显示出显著更低的dCq表达（3.821对4.530；p<0.01）。然而，miRNA水平不受食物摄入的影响。早晨（空腹）和中午（正常进食后）采集的样本之间的差异未显示出任何显著差异。在实验室诊断中，miRNA可被认为是一种相对稳定的数据，但显然每个新的检测方法都需要进行全面评估。健康志愿者中记录的miRNA稳定性显示了它们在寻找肿瘤学创新生物标志物方面的潜力。
