A four-gene methylation marker panel as triage test in high-risk human papillomavirus positive patients
A four-gene methylation marker panel as triage test in high-risk human papillomavirus positive patients
复制标题
四基因甲基化标记物组作为高危人乳头瘤病毒阳性患者的分类测试
DOI：
10.1002/ijc.26326
复制
发表时间：
2012-04-15
影响因子：
6.4
通讯作者：
Wisman, G. B. A.
中科院分区：
文献类型：
--
作者：
Eijsink, J. J. H.;Lendvai, A.;Wisman, G. B. A.
迫切需要具有高灵敏度和特异性的宫颈肿瘤特异性生物标志物，例如DNA甲基化标志物，以改善当前基于人群的宫颈肿瘤（前）恶性筛查。我们的目的是确定新的宫颈肿瘤特异性DNA甲基化标志物，并设计和验证一个甲基化标志物面板，用于高危人乳头瘤病毒（hr-HPV）阳性患者的分诊。首先，在新型OpenArray（TM）平台上对来自84名宫颈癌患者和106名正常宫颈的冷冻组织样本进行高通量定量甲基化特异性PCR（QMSP），该平台代表213个癌症特异性甲基化基因的424个引物。第二，通过LightCycler（R）MSP在来自27名宫颈癌患者和20名正常宫颈和ROC的冷冻组织上验证前20个区别性甲基化标志物，并评估测试特征。三个新的甲基化标记物（JAM 3，EPB 41 L3和TERT），随后与C13 ORF 18在我们的四基因甲基化面板相结合。在第三步中，我们的甲基化组在宫颈刮片中检测到94%（70/74）的宫颈癌，而在第四步中，检测到82%（32/39）的宫颈上皮内瘤变3级或更高（CIN 3+）和65%（44/68）的CIN 2+，其中21%的甲基化组阳性病例。
Cervical neoplasia-specific biomarkers, e.g. DNA methylation markers, with high sensitivity and specificity are urgently needed to improve current population-based screening on (pre)malignant cervical neoplasia. We aimed to identify new cervical neoplasia-specific DNA methylation markers and to design and validate a methylation marker panel for triage of high-risk human papillomavirus (hr-HPV) positive patients. First, high-throughput quantitative methylation-specific PCRs (QMSP) on a novel OpenArray (TM) platform, representing 424 primers of 213 cancer specific methylated genes, were performed on frozen tissue samples from 84 cervical cancer patients and 106 normal cervices. Second, the top 20 discriminating methylation markers were validated by LightCycler (R) MSP on frozen tissue from 27 cervical cancer patients and 20 normal cervices and ROCs and test characteristics were assessed. Three new methylation markers were identified (JAM3, EPB41L3 and TERT), which were subsequently combined with C13ORF18 in our four-gene methylation panel. In a third step, our methylation panel detected in cervical scrapings 94% (70/74) of cervical cancers, while in a fourth step 82% (32/39) cervical intraepithelial neoplasia grade 3 or higher (CIN3+) and 65% (44/68) CIN2+ were detected, with 21% positive cases for