Microfluidic-based multiplex qRT-PCR identifies diagnostic and prognostic microRNA signatures in the sera of prostate cancer patients.
Microfluidic-based multiplex qRT-PCR identifies diagnostic and prognostic microRNA signatures in the sera of prostate cancer patients.
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DOI：
10.1158/0008-5472.can-10-1229
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发表时间：
2011-01-15
期刊：
影响因子：
11.2
通讯作者：
Blelloch R
中科院分区：
文献类型：
--
作者：
Moltzahn F;Olshen AB;Baehner L;Peek A;Fong L;Stöppler H;Simko J;Hilton JF;Carroll P;Blelloch R
最近的前列腺特异性抗原（PSA）为基础的筛选试验表明，迫切需要新的和非侵入性的生物标志物识别策略，以提高前列腺癌的行为预测。血清和血浆中的非编码microRNA（miRNAs）已被证明具有作为生理和病理条件的非侵入性标记物的潜力。为了鉴定诊断前列腺癌和与前列腺癌预后相关的血清miRNAs，我们开发了一种多重定量逆转录PCR（qRT-PCR）方法，该方法包括多重PCR产物的纯化，然后在微流体芯片上进行单重分析以评估384种人类miRNAs。使用Dgcr 8和Dicer敲除（小RNA缺陷）小鼠ES细胞（mESC）作为基准，我们证实了我们的技术的有效性，同时发现了先前公开的方法中显著缺乏准确性。我们分析了来自健康男性和未经治疗的前列腺癌患者的48份血清，这些患者具有不同的CAPRA（前列腺癌风险评估）评分，我们确定了允许诊断癌症患者并与预后相关的miRNA特征。这些血清特征包括致癌和肿瘤抑制miRNA，表明在前列腺癌进展中的功能作用。
Recent prostate specific antigen (PSA) based screening trials indicate an urgent need for novel and non-invasive biomarker identification strategies to improve the prediction of prostate cancer behavior. Non-coding microRNAs (miRNAs) in the serum and plasma have been shown to have potential as non-invasive markers for physiological and pathological conditions. To identify serum miRNAs that diagnose and correlate with prognosis of prostate cancer, we developed a multiplex quantitative reverse transcription PCR (qRT-PCR) method involving purification of multiplex PCR products followed by uniplex analysis on a microfluidics chip to evaluate 384 human miRNAs. Using Dgcr8 and Dicer knockout (small RNA - deficient) mouse ES cells (mESC) as the benchmark, we confirmed the validity of our technique, while uncovering a significant lack of accuracy in previously published methods. Profiling 48 sera from healthy men and untreated prostate cancer patients with differing CAPRA (Cancer of the Prostate Risk Assessment) scores, we identified miRNA signatures that allow to diagnose cancer patients and correlate with prognosis. These serum signatures include oncogenic and tumor suppressive miRNAs suggesting functional roles in prostate cancer progression.