Margue Christiane, Reinsbach Susanne, Philippidou Demetra, Beaume Nicolas, Walters Casandra, Schneider Jochen G, Nashan Dorothée, Behrmann Iris, Kreis Stephanie
Life Sciences Research Unit, University of Luxembourg, Luxembourg.
Klinikum Dortmund GmbH, Germany.
Oncotarget. 2015 May 20;6(14):12110-27. doi: 10.18632/oncotarget.3661.
MiRNAs are increasingly recognized as biomarkers for the diagnosis of cancers where they are profiled from tumor tissue (intracellular miRNAs) or serum/plasma samples (extracellular miRNAs). To improve detection of reliable biomarkers from blood samples, we first compiled a healthy reference miRNome and established a well-controlled analysis pipeline allowing for standardized quantification of circulating miRNAs. Using whole miRNome and custom qPCR arrays, miRNA expression profiles were analyzed in 126 serum, whole blood and tissue samples of healthy volunteers and melanoma patients and in primary melanocyte and keratinocyte cell lines. We found characteristic signatures with excellent prognostic scores only in late stage but not in early stage melanoma patients. Upon comparison of melanoma tissue miRNomes with matching serum samples, several miRNAs were identified to be exclusively tissue-derived (miR-30b-5p, miR-374a-5p and others) while others had higher expression levels in serum (miR-3201 and miR-122-5p). Here we have compiled a healthy and widely applicable miRNome from serum samples and we provide strong evidence that levels of cell-free miRNAs only change significantly at later stages of melanoma progression, which has serious implications for miRNA biomarker studies in cancer.
微小RNA（miRNAs）越来越被认为是癌症诊断的生物标志物，可从肿瘤组织（细胞内miRNAs）或血清/血浆样本（细胞外miRNAs）中对其进行分析。为了提高从血液样本中检测可靠生物标志物的能力，我们首先编制了一份健康参考miRNome，并建立了一个严格控制的分析流程，以实现循环miRNAs的标准化定量。使用全miRNome和定制qPCR阵列，对健康志愿者和黑色素瘤患者的126份血清、全血和组织样本以及原代黑素细胞和角质形成细胞系中的miRNA表达谱进行了分析。我们发现，仅在晚期黑色素瘤患者中发现了具有优异预后评分的特征性信号，而在早期患者中未发现。在将黑色素瘤组织miRNome与匹配的血清样本进行比较时，发现有几种miRNAs是仅来源于组织的（如miR-30b-5p、miR-374a-5p等），而其他一些miRNAs在血清中的表达水平更高（如miR-3201和miR-122-5p）。在这里，我们从血清样本中编制了一份健康且广泛适用的miRNome，并提供了有力证据表明，游离miRNAs的水平仅在黑色素瘤进展的后期才会发生显著变化，这对癌症中的miRNA生物标志物研究具有重要意义。
