Effects of preanalytic variables on circulating microRNAs in whole blood.
Effects of preanalytic variables on circulating microRNAs in whole blood.
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DOI：
10.1158/1055-9965.epi-14-0550
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发表时间：
2014-12
期刊：
影响因子：
--
通讯作者：
Liu S
中科院分区：
文献类型：
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作者：
Zhao H;Shen J;Hu Q;Davis W;Medico L;Wang D;Yan L;Guo Y;Liu B;Qin M;Nesline M;Zhu Q;Yao S;Ambrosone CB;Liu S
过去十年的研究表明，全血循环中的microrna作为癌症检测的生物标志物具有临床潜力。然而，在临床应用已鉴定的循环microrna之前，必须进行以生物标本为重点的研究，以确定可能显著影响循环microrna水平的分析前变量。在本研究中，我们利用Roswell Park癌症研究所的数据库和生物储存库（DBBR）的独特资源，进行了两步分析，以确定全血中的内控循环microrna，然后研究选择的主要分析前变量（即处理延迟、储存条件、储存时间和冷冻/解冻周期）如何影响循环microrna的检测。在第一步的发现阶段，我们确定了三种microrna，包括miR-346、miR-134和miR-934，它们的水平在病例对照组之间以及每组内部的个体间表现出最小的变化。在进一步的验证分析中，验证了miR-346和miR-134的一致性，但没有验证miR-934的一致性。在第二步，使用miR-346和miR-134作为内部对照，我们观察到随着冻融循环次数的增加，miR-346和miR-134的水平都显著降低（P <0.0001），改变其他加工和储存条件不影响miRNA水平。在血浆样品的平行分析中，miR-16的水平随着处理延迟和冻融循环次数的增加而显著降低，但不受储存条件和持续时间的影响。这项研究的结果强调了在临床应用前对循环microrna进行以生物标本为重点的研究的必要性。
Research in the last decade suggests the clinical potential of circulating microRNAs in whole blood as biomarkers for cancer detection. However, before applying the identified circulating microRNAs clinically, biospecimen-focused research has to be performed to identify possible pre-analytic variables which may significantly affect the levels of circulating microRNAs. In this study, utilizing a unique resource of the Data Bank and BioRepository (DBBR) at Roswell Park Cancer Institute, we conducted a two-step analysis to identify internal control circulating microRNAs in whole blood, and then to study how selected major pre-analytic variables (namely, processing delay, storage condition, storage time, and freeze/thaw cycles) might affect the detection of circulating microRNAs. In the discovery phase of the first step, we identified three microRNAs, including miR-346, miR-134 and miR-934, whose levels exhibited the smallest variation between the case-control groups, as well as within each group inter-individually. In the further validation analysis, the consistency was validated for miR-346 and miR-134, but not for miR-934. At the second step, using miR-346 and miR-134 as internal controls, we observed that as numbers of freeze/thaw cycles increased, levels of both miR-346 and miR-134 were significantly decreased (P for trend <0.0001), varying other processing and storage conditions did not affect miRNA levels. In the paralleled analysis in plasma samples, levels of miR-16 were significantly decreased by increasing processing delay and increasing numbers of freeze/thaw cycles, but not affected by storage condition and duration. The results from this study highlight the necessity of biospecimen-focused research on circulating microRNAs before clinical utilization.