Oral squamous cell carcinoma (OSCC) is an aggressive human malignancy. Because of late diagnosis and recurrence of OSCC, the treatment of patients with OSCC is often ineffective. Thus, finding novel biomarkers of OSCC are essential. Here we derived a methylation marker by utilizing methylation microarray data and testing its capacity in cross-sectional study designed for OSCC detection and screening. According to bioinformatics analysis of total of 27,578 cg sites, cg22881914 of Nidogen 2 (NID2) methylation was selected for evaluation. Next, we confirmed the methylation status by bisulfite sequencing from the microdissected OSCC cells in comparison with the microdissected oral epithelia. Subsequently, we developed a simple technique using real-time PCR with the specific probe to examine the ability for the detection of OSCC in the oral epithelial samples, which included 103 oral rinse and 82 oral swab samples. Based on the comparison of microdissected tissue, cg22881914 of NID2 was proved to be methylated in most OSCC cells but unmethylated in the normal oral epithelia. Furthermore, the methylated NID2-relied quantitative PCR approach has demonstrated that this marker assists in distinguishing among patients with OSCC from normal oral epithelia, smokers, and patients with oral lichen planus using the non-invasive oral rinse and swab samples. Specific methylation at cg22881914 of NID2 of OSCC could be used as an important potential marker for detecting OSCC. Thus, to certify the utility of this marker, further studies with a larger sample size are needed.

中文翻译：

---

NID2启动子甲基化用于口腔鳞状细胞癌筛查的评估

口腔鳞状细胞癌（OSCC）是一种侵袭性的人类恶性肿瘤。由于OSCC的诊断和复发较晚，因此OSCC患者的治疗通常无效。因此，寻找新的OSCC生物标志物至关重要。在这里，我们通过利用甲基化微阵列数据并在为OSCC检测和筛选而设计的横断面研究中测试了其能力，得出了甲基化标记。根据对总共27,578 cg位点的生物信息学分析，选择Nidogen 2（NID2）甲基化的cg22881914进行评估。接下来，我们与经显微解剖的口腔上皮细胞相比，通过亚硫酸氢盐测序从显微解剖的OSCC细胞中确认了甲基化状态。后来，我们开发了一种使用实时PCR和特定探针的简单技术，以检测口腔上皮样品中OSCC的检测能力，其中包括103次口腔冲洗液和82次口腔拭子样品。基于显微组织的比较，NID2的cg22881914被证明在大多数OSCC细胞中被甲基化，而在正常口腔上皮细胞中未被甲基化。此外，甲基化的NID2依赖的定量PCR方法已证明，该标记物可使用无创口腔冲洗液和拭子样本帮助区分OSCC患者与正常口腔上皮细胞，吸烟者以及扁平苔藓患者。OSCC NID2的cg22881914处的特异性甲基化可以用作检测OSCC的重要潜在标志物。因此，为了证明此标记的实用性，