Absolute quantification of cell-free microRNAs in cancer patients.
Absolute quantification of cell-free microRNAs in cancer patients.
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DOI：
10.18632/oncotarget.3859
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发表时间：
2015-06-10
期刊：
影响因子：
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通讯作者：
Negrini M
中科院分区：
文献类型：
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作者：
Ferracin M;Lupini L;Salamon I;Saccenti E;Zanzi MV;Rocchi A;Da Ros L;Zagatti B;Musa G;Bassi C;Mangolini A;Cavallesco G;Frassoldati A;Volpato S;Carcoforo P;Hollingsworth AB;Negrini M
使用血液中循环的 microRNA (miRNA) 作为癌症生物标志物的假设是几年前基于有希望的初步结果提出的。然而，在一些令人兴奋的发现之后，很明显，准确定量无细胞 miRNA 比预期更具挑战性。困难与许多（如果不是全部）分析前和分析后变量对最终结果的强烈影响有关。在这项研究中，我们使用目前可用的高通量技术来鉴定乳腺癌、结直肠癌、肺癌、甲状腺和黑色素瘤肿瘤患者以及健康对照者血浆和血清中存在的 miRNA。然后，我们使用液滴数字技术评估了来自健康对照和癌症患者的 207 个样本中九种不同 miRNA（miR-320a、miR-21-5p、miR-378a-3p、miR-181a-5p、miR-3156-5p、miR-2110、miR-125a-5p、miR-425-5p、miR-766-3p）的绝对水平。 PCR（ddPCR）技术。根据组织来源，我们鉴定了在一种或多种癌症类型中特异性调节的 miRNA。使用两个独立队列进一步验证了乳腺癌患者血清中 miR-181a-5p 水平的显着降低，一组来自意大利 (n = 70)，一组来自美国 (n = 90)，AUC 分别为 0.66 和 0.73。这项研究最终支持使用无细胞 miRNA 作为癌症生物标志物，并提出 miR-181a-5p 作为诊断乳腺癌生物标志物。
The hypothesis to use microRNAs (miRNAs) circulating in the blood as cancer biomarkers was formulated some years ago based on promising initial results. After some exciting discoveries, however, it became evident that the accurate quantification of cell-free miRNAs was more challenging than expected. Difficulties were linked to the strong impact that many, if not all, pre- and post- analytical variables have on the final results. In this study, we used currently available high-throughput technologies to identify miRNAs present in plasma and serum of patients with breast, colorectal, lung, thyroid and melanoma tumors, and healthy controls. Then, we assessed the absolute level of nine different miRNAs (miR-320a, miR-21-5p, miR-378a-3p, miR-181a-5p, miR-3156-5p, miR-2110, miR-125a-5p, miR-425-5p, miR-766-3p) in 207 samples from healthy controls and cancer patients using droplet digital PCR (ddPCR) technology. We identified miRNAs specifically modulated in one or more cancer types, according to tissue source. The significant reduction of miR-181a-5p levels in breast cancer patients serum was further validated using two independent cohorts, one from Italy (n = 70) and one from US (n = 90), with AUC 0.66 and 0.73 respectively. This study finally powers the use of cell-free miRNAs as cancer biomarkers and propose miR-181a-5p as a diagnostic breast cancer biomarker.