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尿液样本中的游离DNA完整性分析

Cell-Free DNA Integrity Analysis in Urine Samples.

作者信息

Casadio Valentina, Salvi Samanta, Martignano Filippo, Gunelli Roberta, Ravaioli Sara, Calistri Daniele

机构信息

Biosciences Laboratory, Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS;

Biosciences Laboratory, Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS.

出版信息

J Vis Exp. 2017 Jan 5(119):55049. doi: 10.3791/55049.

Abstract

Although the presence of circulating cell-free DNA in plasma or serum has been widely shown to be a suitable source of biomarkers for many types of cancer, few studies have focused on the potential use of urine cell-free (UCF) DNA. Starting from the hypotheses that normal apoptotic cells produce highly fragmented DNA and that cancer cells release longer DNA, the potential role of UCF DNA integrity was evaluated as an early diagnostic marker capable of distinguishing between patients with prostate or bladder cancer and healthy individuals. A UCF DNA integrity analysis is proposed on the basis of four quantitative real-time PCRs of four sequences longer than 250 bp: c-MYC, BCAS1, HER2, and AR. Sequences that frequently have an increased DNA copy number in bladder and prostate cancers were chosen for the analysis, but the method is flexible, and these genes could be substituted with other genes of interest. The potential utility of UCF DNA as a source of biomarkers has already been demonstrated for urologic malignancies, thus paving the way for further studies on UCF DNA characterization. The UCF DNA integrity test has the advantage of being non-invasive, rapid, and easy to perform, with only a few milliliters of urine needed to carry out the analysis.

摘要

尽管血浆或血清中循环游离DNA的存在已被广泛证明是多种癌症生物标志物的合适来源,但很少有研究关注尿游离(UCF)DNA的潜在用途。基于正常凋亡细胞产生高度片段化DNA以及癌细胞释放更长DNA的假设,评估了UCF DNA完整性作为能够区分前列腺癌或膀胱癌患者与健康个体的早期诊断标志物的潜在作用。基于对四个长度超过250 bp的序列(c-MYC、BCAS1、HER2和AR)进行的四次定量实时PCR,提出了一种UCF DNA完整性分析方法。选择了在膀胱癌和前列腺癌中经常出现DNA拷贝数增加的序列进行分析,但该方法具有灵活性,这些基因可以被其他感兴趣的基因替代。UCF DNA作为生物标志物来源的潜在效用已在泌尿系统恶性肿瘤中得到证实,从而为进一步研究UCF DNA特征铺平了道路。UCF DNA完整性检测具有非侵入性、快速且易于操作的优点,进行分析仅需几毫升尿液。

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