我们最近采用基因组范围的方法鉴定了一些新的甲基化标记物，这些标记物在不同宫颈上皮内瘤变（CIN）分级和宫颈癌中进行了评估。本文中我们通过对组织学确定的结果进行了这些基因的甲基化状态验证，并不考虑人乳头瘤病毒感染情况，并且采用了独立的研究人群。CA10、DPP10、FMN2和HAS1（发现集：54个正常，50个CIN1，40个CIN2，42个CIN3）通过针对亚硫酸盐目标测序（Illumina MiSeq平台）在258个（训练集：100个正常，50个CIN1，50个CIN2，50个CIN3，8个癌症）和373个（验证集：100个正常，57个CIN1，61个CIN2，53个CIN3，102个癌症）由医师收集的样本（PreservCyt）进行了评估。使用来自训练集的针对放大测序数据用于94个正常样本和8个癌症样本，我们计算了每个基因的甲基化值中位数。这些值进行了求和并标准化，以计算一个四基因标记多基因得分（MPS）。我们分别在训练集和验证集中比较了MPS与从正常到CIN分级和癌症的进展之间的关系，并通过接受者操作特征曲线来测试其临床性能。MPS随着CIN分级的增加而增加，并且在训练（曲线下面积，AUC=0.9950）和验证（AUC=0.9337）集中准确预测了宫颈癌，将正常与癌症进行对比。在100％特异性最高阈值下，宫颈癌的检测敏感性为67.7％；而将特异性降低至95％则将敏感性提高到84.3％。有必要在前瞻性研究中进一步评估这些生物标志物。©2023 年作者。国际癌症杂志由约翰•威立出版社代表国际癌症合作组织出版。

We have recently identified, using a genome-wide approach, new methylation markers which were evaluated among various cervical intraepithelial neoplasia (CIN) grades and cervical cancer. We herein validate the methylated state of these genes in independent study populations, based on histology ascertained outcomes regardless of human papillomavirus status. CA10, DPP10, FMN2 and HAS1 (discovery set: 54 normal, 50 CIN1, 40 CIN2, 42 CIN3) were evaluated by targeted bisulfite next generation sequencing (NGS) (Illumina MiSeq platform) in 258 (training set: 100 normal, 50 CIN1, 50 CIN2, 50 CIN3, 8 cancers) and 373 (validation set: 100 normal, 57 CIN1, 61 CIN2, 53 CIN3, 102 cancers) physician-collected samples (PreservCyt). Using targeted amplification NGS data from the training set for 94 normal and eight cancer samples, we calculated for each gene the median methylation value. These were summed and normalized to compute a four-gene Marker Polygenic Score (MPS). We compared the relationship between MPS and progression from normal through CIN grades and cancer, separately in the training and validation sets, and tested its clinical performance via receiver-operating characteristic curves. MPS increased with increasing CIN grade, and accurately predicted cervical cancer in the training (area under the curve, AUC = 0.9950) and validation (AUC = 0.9337) sets, comparing normal to cancer. Using the highest threshold of 100% specificity, sensitivity for detection of cervical cancer was 67.7%; whereas reducing specificity to 95% increased sensitivity to 84.3%. Further evaluation of these biomarkers is warranted in prospective studies.© 2023 The Authors. International Journal of Cancer published by John Wiley & Sons Ltd on behalf of UICC.