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  • Title: Limited Role of Promoter Methylation of MGMT and C13ORF18 in Triage of Low-Grade Squamous Intraepithelial Lesion.
    Author: Sun LL, Liu Y, Sun X, Pan L, Wu D, Wang YD.
    Journal: Chin Med J (Engl); 2018 Apr 20; 131(8):939-944. PubMed ID: 29664054.
    Abstract:
    BACKGROUND: Promoter methylation of MGMT and C13ORF18 has been confirmed as a potential biomarker for early diagnosis of cervical cancer. The aim of this study was to evaluate the performance of MGMT and C13ORF18 promoter methylation for triage of cytology screening samples and explore the potential mechanism. METHODS: Methylation-sensitive high-resolution melting was used to detect promoter methylation of MGMT and C13ORF18 in 124 cervical samples. High-risk human papillomavirus (HR-HPV) was detected by the Digene Hybrid Capture 2®. Gene methylation frequencies in relation to cervical intraepithelial neoplasia (CIN) were analyzed. Frequencies were compared by Chi-square tests. The expression of gene biomarkers and methylation regulators was analyzed by immunohistochemical staining, real-time fluorescence quantitative polymerase chain reaction, and Western blot. RESULTS: For triage of low-grade squamous intraepithelial lesion (LSIL), gene methylation increased specificity from 4.0% of HR-HPV detection to 30.8% of MGMT (χ2 = 9.873, P = 0.002) and to 50.0% of C13ORF18 (χ2 = 21.814, P = 0.001). For triage of atypical squamous cells of undetermined significance, HR-HPV detection had higher positive predictive value of 54.8%. Either MGMT or C13ORF18 methylation combined with HR-HPV increased the negative predictive value to 100.0% (χ2 = 9.757, P = 0.002). There was no relationship between MGMT and C13ORF18 expression and DNA methylation (χ2 = 0.776, P = 0.379 and χ2 = 1.411, P = 0.235, respectively). MBD2 protein level in cervical cancer was relatively lower than normal cervical tissue (t = 4.11, P = 0.006). CONCLUSIONS: HR-HPV detection is the cornerstone for triage setting of CIN. Promoter methylation of MGMT and C13ORF18 plays a limited role in triage of LSIL. Promoter methylation of both genes may not be the causes of gene silence. MGMT和C13ORF18基因甲基化对分流低级别宫颈上皮内病变起有限作用摘要研究背景：MGMT和 C13ORF18基因甲基化是宫颈癌早期诊断潜在的分子标记。本研究目的是评价MGMT和 C13ORF18基因甲基化对宫颈细胞学筛查样本的分流作用及机制探讨。 材料与方法：应用甲基化敏感性高通量溶解曲线法（MS-HRM）对124例宫颈脱落细胞学样本行MGMT 和C13ORF18启动子甲基化检测，采用HC2法检测高危型HPV感染，分析基因甲基化与各级宫颈上皮内瘤变的关系。采用卡方检验进行率的比较。采用蛋白印迹法、RT-PCR及免疫组化法检测MGMT、C13ORF18以及甲基化调节基因的表达。 结果：对低级别宫颈上皮内病变（low-grade squamous intraepithelial lesion，LSIL）的分流，高危HPV检测的特异性为4.0%，MGMT甲基化将诊断高级别病变的特异性提高至30.8% (X2 = 9.873, P = 0.002)，C13ORF18甲基化将特异性提高至50.0% (X2 = 21.814, P = 0.001). 对不能明确意义的非典型鳞状细胞（atypical squamous cells of undetermined significance，ASCUS）的分流，高危HPV检测具有较高的阳性预测值（54.8%），MGMT或C13ORF18甲基化联合高危HPV检测均可将阴性预测值显著提高至100.0% (X2 = 9.757, P = 0.002)。MGMT和C13ORF18甲基化与该基因表达无关(X2 = 0.776, P = 0.379；X2 = 1.411, P = 0.235)。宫颈癌组织MBD2蛋白表达显著低于正常宫颈组织 (t = 4.11, P = 0.006)。 结论：高危HPV检测是宫颈病变分流的基石，MGMT和C13ORF18启动子甲基化对分流低级别宫颈上皮内病变起有限作用。MGMT和C13ORF18甲基化可能不是基因沉默的原因。.
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