MicroRNA Stability in Postmortem FFPE Tissues: Quantitative Analysis Using Autoptic Samples from Acute Myocardial Infarction Patients.
MicroRNA Stability in Postmortem FFPE Tissues: Quantitative Analysis Using Autoptic Samples from Acute Myocardial Infarction Patients.
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DOI：
10.1371/journal.pone.0129338
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Osawa M
中科院分区：
文献类型：
--
作者：
Kakimoto Y;Kamiguchi H;Ochiai E;Satoh F;Osawa M
MicroRNAs(MiRNAs)是一种非常短(18-24个核苷酸)的核酸，在许多生物组织中表达，已被证明比较长的RNA分子(如mRNAs)更能抵抗极端温度和pH。由于miRNAs参与不同的生物学过程，并对各种细胞应激做出反应，它们作为诊断生物标志物和/或治疗靶点的有效性最近得到了探索。在这里，我们评估了miRNA定量在急性心肌梗死(AMI)患者死后心脏组织检查中的有效性。在患者死亡的一周内收集心脏组织，并将其冷冻(19个样本)或在福尔马林中固定长达三年(36个样本)。用电泳图评估RNA的完整性，在长期固定的样本中，死亡后似乎有较长的RNA片断。我们还对7个miRNA和3个其他小RNA进行了定量聚合酶链式反应，以便为我们的尸检分析确定适当的对照。我们的数据表明miR-191和miR-26b比其他类型的小RNA分子更适合，因为它们在死后和长期固定后都能稳定地被检测到。此外，我们还应用我们的定量方法，使用这些内源性对照，评估了三个先前识别的miRNA生物标记物miR-1、miR-208b和miR-499a在急性心肌梗死患者福尔马林固定组织中的表达。虽然与对照组相比，急性心肌梗死患者的miR-1和miR-208B分别减少(1.4倍)和增加(1.2倍)，但这些变化的意义受到样本量的限制。相反，在急性心肌梗死患者中miR-499a的相对水平显著降低(2.1倍)。这项研究强调了miRNAs在死亡和长期固定后的稳定性，验证了它们在尸检中作为急性心肌梗死的可靠生物标志物的使用。
MicroRNAs (miRNAs) are very short (18–24 nucleotides) nucleic acids that are expressed in a number of biological tissues and have been shown to be more resistant to extreme temperatures and pH compared to longer RNA molecules, like mRNAs. As miRNAs contribute to diverse biological process and respond to various kinds of cellular stress, their utility as diagnostic biomarkers and/or therapeutic targets has recently been explored. Here, we have evaluated the usefulness of miRNA quantification during postmortem examination of cardiac tissue from acute myocardial infarction (AMI) patients. Cardiac tissue was collected within one week of the patient’s death and either frozen (19 samples) or fixed in formalin for up to three years (36 samples). RNA integrity was evaluated with an electropherogram, and it appears that longer RNAs are fragmented after death in the long-term fixed samples. Quantitative PCR was also performed for seven miRNAs and three other small RNAs in order to determine the appropriate controls for our postmortem analysis. Our data indicate that miR-191 and miR-26b are more suitable than the other types of small RNA molecules as they are stably detected after death and long-term fixation. Further, we also applied our quantitation method, using these endogenous controls, to evaluate the expression of three previously identified miRNA biomarkers, miR-1, miR-208b, and miR-499a, in formalin-fixed tissues from AMI patients. Although miR-1 and miR-208b decreased (1.4-fold) and increased (1.2-fold), respectively, in the AMI samples compared to the controls, the significance of these changes was limited by our sample size. In contrast, the relative level of miR-499a was significantly decreased in the AMI samples (2.1-fold). This study highlights the stability of miRNAs after death and long-term fixation, validating their use as reliable biomarkers for AMI during postmortem examination.
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影响因子：
14.9
作者：
Imbeaud S;Graudens E;Boulanger V;Barlet X;Zaborski P;Eveno E;Mueller O;Schroeder A;Auffray C
通讯作者：
Auffray C
DOI：
10.1097/paf.0b013e31828879cd
发表时间：
2013-09-01
影响因子：
1
作者：
Kakimoto, Yu;Tsuruyama, Tatsuaki;Tamaki, Keiji
通讯作者：
Tamaki, Keiji
影响因子：
4.8
作者：
Becker, C.;Hammerle-Fickinger, A.;Pfaffl, M. W.
通讯作者：
Pfaffl, M. W.
影响因子：
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作者：
Corsten, Maarten F.;Dennert, Robert;Schroen, Blanche
通讯作者：
Schroen, Blanche
影响因子：
3.7
作者：
Meng W;McElroy JP;Volinia S;Palatini J;Warner S;Ayers LW;Palanichamy K;Chakravarti A;Lautenschlaeger T
通讯作者：
Lautenschlaeger T