Methylation of CADM1 and MAL together with HPV status in cytological cervical specimens serves an important role in the progression of cervical intraepithelial neoplasia
Methylation of CADM1 and MAL together with HPV status in cytological cervical specimens serves an important role in the progression of cervical intraepithelial neoplasia
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DOI：
10.3892/ol.2018.9505
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发表时间：
2018-12-01
期刊：
影响因子：
2.9
通讯作者：
Danko, Jan
中科院分区：
文献类型：
--
作者：
Mersakova, Sandra;Holubekova, Veronika;Danko, Jan
宫颈癌是影响女性人口的第二种最常见的癌症类型。宫颈鳞癌的发展需要数年时间，并涉及到称为宫颈上皮内瘤变(CIN)的癌前阶段。疾病发展中的一个关键因素是人类乳头瘤病毒(HPV)感染，它启动了癌症的发生。此外，CC还受到表观遗传变化的影响，如DNA甲基化，导致某些基因的激活或排除，以及启动子中胞嘧啶的超甲基化，从而关闭先前活跃的基因。大多数DNA甲基化事件发生在胞嘧啶-鸟嘌呤核苷酸上，在人类基因组中被称为CpG岛。本研究的目的是利用细胞学标本检测两个抑癌基因细胞黏附分子1(CADM1)和T淋巴细胞成熟相关蛋白(MAL)内含子序列的甲基化水平，并通过焦磷酸测序寻找与CIN相关的潜在生物标志物。从CIN患者的宫颈涂片中提取DNA，并以健康患者为对照组。样本通过亚硫酸氢钠处理和随后的焦磷酸测序进行转换，以检测选定基因的甲基化状态。通过聚合酶链式反应分析每个样本中是否存在HPVDNA感染。在总样本数(n=91)中，本研究证实39例(42.85%)存在一种或两种高危HPV亚型，HPV感染与CIN2+病变显著相关。与HPVDNA阴性组相比，HPVDNA阳性组的Mal和CADM1基因甲基化水平显著升高[P=0.0097，95%可信区间(CI)：(-0.030，-0.003)/P=0.0024，95%CI：(-0.06，-0.01)]，且甲基化水平随宫颈病变程度的加重而增加。本研究使用逻辑回归来模拟病例/对照状态(对照组与DG)之间的依赖关系。1-4)。MAL曲线下面积分别为：宫颈炎症84%，CIN1 71%，CIN2+73.4%，鳞癌77%，CADM1：宫颈炎症88.6%，CIN1 68%，CIN2+80%，鳞癌89%。本研究证实，在HPV16/18阳性的患者中，个体CPGS的甲基化水平差异有统计学意义，且显著高于HPV16/18阳性患者的中位甲基化水平。CADM1在CIN过渡期间几乎每个研究组的甲基化水平都高于MAL，有望成为未来研究的生物标志物。
Cervical cancer (CC) is the second most common type of cancer affecting the female population. The development of CC takes several years, and involves a precancerous stage known as cervical intraepithelial neoplasia (CIN). A key factor in the development of disease is the human papillomavirus (HPV) infection, which initiates carcinogenesis. Furthermore, CC is also impacted by epigenetic changes such as DNA methylation, which causes activation or exclusion of certain genes, and the hypermethylation of cytosines in promoters, thereby switching off previously active genes. The majority of DNA methylation events occur at cytosine-guanine nucleotides, which in the human genome are known as CpG islands. The aim of the present study was to investigate the methylation levels in intronic sequences of the two tumor suppressor genes cell adhesion molecule 1 (CADM1) and T-lymphocyte maturation associated protein (MAL) using cytological samples and to identify potential biomarkers involved in CIN by pyrosequencing. DNA was isolated from cervical smears from patients with CINs, with healthy patients serving as a control group. Samples were converted by treatment with sodium bisulfite and subsequent pyrosequencing to detect the methylation status of the selected genes. The presence of HPV DNA infection analyzed by the polymerase chain reaction, was detected in each sample. Of the total number of samples (n=91), the present study confirmed the presence of one or two high-risk subtypes of HPV in 39 cases (42.85%) and HPV infection was significantly associated with CIN2+ lesions. For the two genes (MAL and CADM1) the present study confirmed that the median methylation was significantly higher in HPV positive patients [P=0.0097, 95% confidence interval (CI): (-0.030, -0.003)/P=0.0024, 95% CI: (-0.06, -0.01)] when compared with patients negative for HPV DNA infection, and the average methylation was demonstrated to be increased with the degree of cervical lesion. The present study used logistical regression to model the dependence between the case/control statuses (control group vs. Dg. 1-4). The area under the curve values for MAL were: 84% for cervical inflammation, 71% for CIN1, 73.4% for CIN2+ and 77% for squamous cell carcinoma (SCC); and for CADM1 were: 88.6% for cervical inflammation, 68% for CIN1, 80% for CIN2+ and 89% for SCC. The present study confirmed that there were statistically significant differences between the methylation levels of individual CpGs and significantly higher median methylation in patients positive for HPV16/18. CADM1 exhibited higher levels of methylation in almost every study group when compared with MAL during the transition of CIN and appeared to be a promising biomarker for future study.