Department of Gastroenterology, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan.
Precursory Research for Embryonic Science and Technology (PRESTO), Japan Science and Technology Agency (JST), Kawaguchi, Saitama, Japan.
JCI Insight. 2016 Jun 2;1(8):e86646. doi: 10.1172/jci.insight.86646.
Pancreatic ductal adenocarcinoma (Pdac) is a malignancy with a poor prognosis due to difficulties in early detection. Although promising biomarkers are increasingly reported, such methods are not yet easy to apply clinically, mainly due to their low reproducibility or technical difficulties. In this study, we developed a convenient and sensitive method for quantifying aberrantly expressed satellite repeat RNAs in sera, which can be used to efficiently detect patients with Pdac. Here, we introduce a Tandem Repeat Amplification by nuclease Protection (TRAP) method combined with droplet digital PCR (ddPCR) to detect human satellite II (HSATII) RNAs, which are specifically expressed in human Pdacs at greater levels than normal tissues but are difficult to measure due to their repetitive sequences and irregularities. HSATII RNA core sequence levels in sera were significantly higher in Pdac patients compared with noncancer patients (median copy number: 14.75 and 3.17 per μl in the training set and 17.35 and 2.9 in the validation set, respectively). In addition, patients with intraductal papillary mucinous neoplasm (IPMN), a precancerous lesion of Pdac, could also be efficiently detected. This method can be routinely applied to screen patients with Pdac and high-risk patients, facilitating the development of preventive medicine for this disease.
胰腺导管腺癌 (Pdac) 是一种预后不良的恶性肿瘤,主要是由于早期检测困难。尽管越来越多有前途的生物标志物被报道,但这些方法尚未易于临床应用,主要是由于其低重现性或技术困难。在这项研究中,我们开发了一种方便且灵敏的方法来定量血清中异常表达的卫星重复 RNA,该方法可用于有效检测 Pdac 患者。在这里,我们介绍了一种串联重复扩增通过核酸酶保护 (TRAP) 方法与液滴数字 PCR (ddPCR) 相结合,以检测人类卫星 II (HSATII) RNA,其在人 Pdacs 中的表达水平高于正常组织,但由于其重复序列和不规则性而难以测量。Pdac 患者血清中的 HSATII RNA 核心序列水平明显高于非癌症患者(训练集的中位数拷贝数分别为 14.75 和 3.17 per μl,验证集分别为 17.35 和 2.9)。此外,还可以有效地检测到胰腺导管内乳头状黏液性肿瘤 (IPMN) 患者,这是 Pdac 的癌前病变。该方法可常规用于筛选 Pdac 患者和高危患者,有利于该疾病的预防医学发展。