肺癌中的液体分析——支气管灌洗中细胞外微小RNA的定量分析

Liquid Profiling in Lung Cancer - Quantification of Extracellular miRNAs in Bronchial Lavage.

作者信息

Schmidt Bernd, Rehbein Grit, Fleischhacker Michael

机构信息

Universitätsklinikum Halle (Saale), Klinik für Innere Medizin, Schwerpunkt Pneumologie, Ernst Grube Str 40, 06120, Halle/Saale, Germany.

Klinik für Innere Medizin - Schwerpunkt Pneumologie und Schlafmedizin DRK Kliniken Berlin∣Mitte, Berlin, Germany.

出版信息

Adv Exp Med Biol. 2016;924:33-37. doi: 10.1007/978-3-319-42044-8_7.

DOI:10.1007/978-3-319-42044-8_7

PMID:27753015

Abstract

Extracellular miRNAs cannot only be isolated from different body fluids like plasma and serum, but also from bronchial lavage samples (BL) obtained by bronchoscopy. Alterations in the expression of microRNAs might be useful for a discrimination of lung cancer patients from patients with a benign lung disease. We profiled extracellular microRNAs from three BL pools of lung cancer patients and three BL pools from a control group (patients with a benign lung disease) with TaqMan MicroRNA Array cards. For the confirmation of these results, we analyzed a panel of eight miRNAs in a qRT-PCR of the BL of 30 different lung cancer and non-cancerous patients. For the data normalization, we used exogenously added cel-miR-39 RNA. Using microRNA arrays, we found a panel of eight microRNAs (hsa-miR 19b-1, 1285, 1289, 1303, 217, 29a-5p, 548-3p, 650) that were differentially expressed between the lung cancer and the non-cancerous group. Further investigation by qPCR revealed five microRNAs (U6 snRNA, hsa-miR 1285, 1303, 29a-5p, 650) that were significantly up-regulated in patients with lung cancer. In bronchial lavage samples, the five microRNAs identified in this study may have a diagnostic potential to be used as biomarkers in lung cancer.

摘要

细胞外微小RNA不仅可以从血浆和血清等不同体液中分离出来，还可以从通过支气管镜检查获得的支气管灌洗样本（BL）中分离出来。微小RNA表达的改变可能有助于区分肺癌患者和良性肺病患者。我们使用TaqMan微小RNA阵列芯片对肺癌患者的三个支气管灌洗样本池和对照组（良性肺病患者）的三个支气管灌洗样本池中的细胞外微小RNA进行了分析。为了证实这些结果，我们在30名不同肺癌患者和非癌症患者的支气管灌洗样本的定量逆转录聚合酶链反应（qRT-PCR）中分析了一组8种微小RNA。为了进行数据标准化，我们使用了外源添加的cel-miR-39 RNA。使用微小RNA阵列，我们发现一组8种微小RNA（hsa-miR 19b-1、1285、1289、1303、217、29a-5p、548-3p、650）在肺癌组和非癌症组之间存在差异表达。通过定量聚合酶链反应（qPCR）进一步研究发现，5种微小RNA（U6小核RNA、hsa-miR 1285、1303、29a-5p、650）在肺癌患者中显著上调。在支气管灌洗样本中，本研究鉴定出的这5种微小RNA可能具有作为肺癌生物标志物的诊断潜力。