van Agthoven Ton, Looijenga Leendert H J
Department of Pathology, Josephine Nefkens Building, Erasmus MC Cancer Institute, Rotterdam, The Netherlands.
Oncotarget. 2016 Jul 27;8(35):58037-58049. doi: 10.18632/oncotarget.10867. eCollection 2017 Aug 29.
Multiple studies, including various methods and overall limited numbers of mostly heterogeneous cases, indicate that the level of embryonic stem cell microRNAs (miRs) (e.g. 371a-3p, 372-3p, 373-3p, and 367-3p) are increased in serum at primary diagnosis of almost all testicular germ cell cancer (TGCC). Here we determine the status of three of these miRs in serum samples of 250 TGCC patients, collected at time of primary diagnosis, compared with 60 non-TGCC patients and 104 male healthy donors. The levels of miRs were measured by the robust ampTSmiR test, including magnetic bead-based miR isolation and target specific pre-amplification followed by real-time quantitative PCR (RT-qPCR) detection. Calibration is performed based on the non-human spike-in ath-miR-159a, and normalization on the endogenous control miR-30b-5p. The serum levels of miR-371a-3p, 373-3p, and 367-3p are informative to accurately detect TGCC patients, both seminomas and non-seminomas, at the time of primary diagnosis (< 0.000). Receiver Operating Characteristic (ROC) analysis demonstrate that the Area Under the Curve (AUC) for miR-371a-3p is 0.951 (being 0.888 for miR-373-3p and 0.861 for miR-367-3p), with a sensitivity of 90%, and a specificity of 86% (positive predictive value of 94% and negative predictive value of 79%). Inclusion of miR-373-3p and 367-3p resulted in a AUC of 0.962, with a 90% sensitivity and 91% specificity. Similar results were obtained using the raw Ct data. Importantly, the results demonstrate that ampTSmiR is not suitable to detect pure teratoma as well as the precursor of TGCC, i.e., Germ Cell Neoplasia (GCNIS). The largest series evaluated so far, demonstrate that detection of the embryonic stem cell miR-371a-3p, 373-3p and 367-3p is highly informative to diagnose patients with a primary TGCC.
多项研究，包括采用各种方法且大多为异质性病例的总体数量有限的研究，表明在几乎所有睾丸生殖细胞癌（TGCC）初诊时，血清中胚胎干细胞微小RNA（miR）（如371a - 3p、372 - 3p、373 - 3p和367 - 3p）的水平会升高。在此，我们测定了250例TGCC患者初诊时采集的血清样本中这三种miR的状态，并与60例非TGCC患者和104名男性健康供体进行比较。通过稳健的ampTSmiR检测法测量miR水平，该方法包括基于磁珠的miR分离、靶标特异性预扩增，随后进行实时定量聚合酶链反应（RT - qPCR）检测。校准基于非人加标对照ath - miR - 159a进行，以内源性对照miR - 30b - 5p进行标准化。miR - 371a - 3p、373 - 3p和367 - 3p的血清水平有助于在初诊时准确检测TGCC患者，包括精原细胞瘤和非精原细胞瘤患者（<0.000）。受试者工作特征（ROC）分析表明，miR - 371a - 3p的曲线下面积（AUC）为0.951（miR - 373 - 3p为0.888，miR - 367 - 3p为0.861），灵敏度为90%，特异性为86%（阳性预测值为94%，阴性预测值为79%）。纳入miR - 373 - 3p和367 - 3p后，AUC为0.962，灵敏度为90%，特异性为91%。使用原始Ct数据也获得了类似结果。重要的是，结果表明ampTSmiR不适用于检测纯畸胎瘤以及TGCC的前驱病变，即生殖细胞肿瘤未成熟型（GCNIS）。迄今为止评估的最大样本系列表明，检测胚胎干细胞miR - 371a - 3p、373 - 3p和367 - 3p对诊断原发性TGCC患者具有很高的参考价值。
