Moltzahn Felix, Olshen Adam B, Baehner Lauren, Peek Andrew, Fong Lawrence, Stöppler Hubert, Simko Jeffry, Hilton Joan F, Carroll Peter, Blelloch Robert
The Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, Department of Urology, UCSF-Helen Diller Family Comprehensive Cancer Center, University of California San Francisco, San Francisco, California 94143, USA.
Cancer Res. 2011 Jan 15;71(2):550-60. doi: 10.1158/0008-5472.CAN-10-1229. Epub 2010 Nov 22.
Recent prostate-specific antigen-based screening trials indicate an urgent need for novel and noninvasive biomarker identification strategies to improve the prediction of prostate cancer behavior. Noncoding microRNAs (miRNA) in the serum and plasma have been shown to have potential as noninvasive markers for physiologic and pathologic conditions. To identify serum miRNAs that diagnose and correlate with the prognosis of prostate cancer, we developed a multiplex quantitative reverse transcription PCR method involving the 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 as the benchmark, we confirmed the validity of our technique and uncovered a considerable lack of accuracy in previously published methods. Profiling 48 sera from healthy men and untreated prostate cancer patients with differing CAPRA scores, we identified miRNA signatures that allow us to diagnose cancer patients and correlate with a prognosis. These serum signatures include oncogenic and tumor-suppressive miRNAs, suggesting functional roles in prostate cancer progression.
近期基于前列腺特异性抗原的筛查试验表明，迫切需要新的非侵入性生物标志物识别策略，以改善对前列腺癌行为的预测。血清和血浆中的非编码微小RNA（miRNA）已显示出作为生理和病理状况非侵入性标志物的潜力。为了鉴定可诊断前列腺癌并与预后相关的血清miRNA，我们开发了一种多重定量逆转录PCR方法，该方法包括纯化多重PCR产物，然后在微流控芯片上进行单重分析，以评估384种人类miRNA。以Dgcr8和Dicer基因敲除（小RNA缺陷型）小鼠胚胎干细胞作为基准，我们证实了该技术的有效性，并发现先前发表的方法存在相当大的准确性不足。对48名健康男性和不同CAPRA评分的未经治疗的前列腺癌患者的血清进行分析，我们鉴定出了能够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抑癌miRNA，提示其在前列腺癌进展中的功能作用。
