Diagnostic and prognostic microRNAs in the serum of breast cancer patients measured by droplet digital PCR.
Diagnostic and prognostic microRNAs in the serum of breast cancer patients measured by droplet digital PCR.
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DOI：
10.1186/s40364-015-0037-0
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发表时间：
2015
影响因子：
11.1
通讯作者：
Negrini M
中科院分区：
文献类型：
--
作者：
Mangolini A;Ferracin M;Zanzi MV;Saccenti E;Ebnaof SO;Poma VV;Sanz JM;Passaro A;Pedriali M;Frassoldati A;Querzoli P;Sabbioni S;Carcoforo P;Hollingsworth A;Negrini M
乳腺癌循环生物标志物包括癌胚抗原和糖类抗原15-3，用于患者随访。由于敏感性和特异性较低，因此需要新的和更有用的生物标志物。血清或血浆中循环稳定的microRNAs(MiRNAs)的存在表明这些微小的RNAs作为癌症生物标记物具有很好的作用。为了获得循环中miRNAs的绝对浓度，并减少分析前和分析变量的影响，我们使用了液滴数字聚合酶链式反应(DdPCR)技术。我们调查了两个独立的乳腺癌患者和无疾病对照人群血清中的五个miRNAs。研究表明，两组乳腺癌患者血清miR-148B-3p和miR-652-3p水平均显著低于对照组。对于这两个miRNAs，通过接收器操作特征曲线分析证实了乳腺癌患者与对照组之间的分层。此外，我们还发现，血清miR-10b-5p水平升高与预后不良的临床生物学指标有关。这项研究揭示了ddPCR方法在定量循环miRNAs方面的有效性。DdPCR定量方法的使用显示了两个独立队列之间在可比较的miRNA绝对浓度方面的非常好的一致性，以及乳腺癌患者与对照组之间调节失调的一致趋势。总体而言，这项研究支持使用定量ddPCR方法来监测诊断和预后肿瘤特异性循环miRNAs的绝对水平。本文的在线版本(doi：10.1186/s40364-0150037-0)包含补充材料，授权用户可以使用。
Breast cancer circulating biomarkers include carcinoembryonic antigen and carbohydrate antigen 15–3, which are used for patient follow-up. Since sensitivity and specificity are low, novel and more useful biomarkers are needed. The presence of stable circulating microRNAs (miRNAs) in serum or plasma suggested a promising role for these tiny RNAs as cancer biomarkers. To acquire an absolute concentration of circulating miRNAs and reduce the impact of preanalytical and analytical variables, we used the droplet digital PCR (ddPCR) technique. We investigated a panel of five miRNAs in the sera of two independent cohorts of breast cancer patients and disease-free controls. The study showed that miR-148b-3p and miR-652-3p levels were significantly lower in the serum of breast cancer patients than that in controls in both cohorts. For these two miRNAs, the stratification of breast cancer patients versus controls was confirmed by receiver operating characteristic curve analyses. In addition, we showed that higher levels of serum miR-10b-5p were associated with clinicobiological markers of poor prognosis. The study revealed the usefulness of the ddPCR approach for the quantification of circulating miRNAs. The use of the ddPCR quantitative approach revealed very good agreement between two independent cohorts in terms of comparable absolute miRNA concentrations and consistent trends of dysregulation in breast cancer patients versus controls. Overall, this study supports the use of the quantitative ddPCR approach for monitoring the absolute levels of diagnostic and prognostic tumor-specific circulating miRNAs. The online version of this article (doi:10.1186/s40364-015-0037-0) contains supplementary material, which is available to authorized users.