Division of Cancer Epidemiology, McGill University, Montréal, QC, Canada.
Gerald Bronfman Department of Oncology, McGill University, Montréal, QC, Canada.
Int J Cancer. 2020 Sep 1;147(5):1264-1274. doi: 10.1002/ijc.32880. Epub 2020 Feb 4.
DNA methylation analysis may improve risk stratification in cervical screening. We used a pan-epigenomic approach to identify new methylation markers along the continuum of cervical intraepithelial neoplasia (CIN) to cervical cancer. Physician-collected samples (54 normal, 50 CIN1, 40 CIN2 and 42 CIN3) were randomly selected from women at a single-center colposcopy clinic. Extracted DNA was subjected to Illumina Infinium EPIC array analysis, and methylation was assessed blinded to histopathological and clinical data. CpG sites whose state of methylation correlated with lesion grade were assessed (Spearman correlation), and a weighted methylation score was calculated comparing normal to CIN3. Validation of the top selected genes was performed in an independent cohort (100 normal, 50 CIN1, 50 CIN2, 50 CIN3 and 8 cervical cancers) of new patients, referred for colposcopic examination at three hospitals, using targeted DNA methylation Illumina amplicon sequencing. The relationship between a combined weighted marker score and progression from normal through precancerous lesions and cervical cancer was compared using one-way ANOVA. Our analyses revealed 7,715 CpGs whose methylation level correlated with progression (from normal to CIN1, CIN2 and CIN3), with a significant trend of increased methylation with lesion grade. We shortlisted a bigenic (hyaluronan synthase 1, HAS1 and ATPase phospholipid transporting 10A, ATP10A corresponding to cg03419058 and cg13944175 sites) marker set; r = 0.55, p < 0.0001. Validation of the four most discriminating genes (CA10, DPP10, FMN2 and HAS1) showed a significant correlation between methylation levels and disease progression (p-value < 2.2 × 10 , adjusted R = 0.952). Translational research of the identified genes to future clinical applications is warranted.
DNA 甲基化分析可能会改善宫颈癌筛查的风险分层。我们使用泛表观基因组学方法来识别沿着宫颈上皮内瘤变（CIN）到宫颈癌连续体的新甲基化标记物。从一家单中心阴道镜诊所的女性中随机选择了医生采集的样本（54 个正常，50 个 CIN1，40 个 CIN2 和 42 个 CIN3）。提取的 DNA 进行了 Illumina Infinium EPIC 阵列分析，并且在不了解组织病理学和临床数据的情况下评估了甲基化状态。评估了与病变等级相关的甲基化 CpG 位点（Spearman 相关性），并计算了比较正常与 CIN3 的加权甲基化评分。使用针对三个医院的阴道镜检查转诊的新患者的独立队列（100 个正常，50 个 CIN1，50 个 CIN2，50 个 CIN3 和 8 个宫颈癌），通过靶向 DNA 甲基化 Illumina 扩增子测序对顶级候选基因进行了验证。使用单向方差分析比较了组合加权标记评分与从正常到癌前病变和宫颈癌的进展之间的关系。我们的分析显示了 7715 个 CpG 位点，其甲基化水平与进展相关（从正常到 CIN1、CIN2 和 CIN3），并且随着病变等级的增加而呈现出明显的甲基化趋势。我们列出了一个双基因（透明质酸合酶 1，HAS1 和 ATP 酶磷脂转运蛋白 10A，ATP10A 对应于 cg03419058 和 cg13944175 位点）标记物集；r = 0.55，p < 0.0001。对四个最具鉴别力的基因（CA10、DPP10、FMN2 和 HAS1）的验证表明，甲基化水平与疾病进展之间存在显著相关性（p 值 < 2.2 × 10 -16 ，调整 R = 0.952）。需要对鉴定出的基因进行转化研究，以将其应用于未来的临床实践。
