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评价 NID2 启动子甲基化用于口腔鳞状细胞癌的筛查。

Evaluation of NID2 promoter methylation for screening of Oral squamous cell carcinoma.

机构信息

Faculty of Medicine, King Mongkut's Institute of Technology Ladkrabang, Bangkok, 10520, Thailand.

Center of Excellence in Molecular Genetics of Cancer and Human Diseases, Department of Anatomy, Faculty of Medicine, Chulalongkorn University, Bangkok, 10330, Thailand.

出版信息

BMC Cancer. 2020 Mar 14;20(1):218. doi: 10.1186/s12885-020-6692-z.

Abstract

BACKGROUND

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.

METHODS

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.

RESULTS

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.

CONCLUSIONS

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.

摘要

背景

口腔鳞状细胞癌(OSCC)是一种侵袭性的人类恶性肿瘤。由于 OSCC 的诊断较晚和复发,OSCC 患者的治疗往往效果不佳。因此,寻找 OSCC 的新型生物标志物至关重要。在这里,我们利用甲基化微阵列数据得出了一个甲基化标志物,并在针对 OSCC 检测和筛查的横断面研究中测试了其能力。

方法

根据总共 27578 个 cg 位点的生物信息学分析,选择 Nidogen 2(NID2)的 cg22881914 进行评估。接下来,我们通过亚硫酸氢盐测序从微切割的 OSCC 细胞中确认了甲基化状态,并与微切割的口腔上皮细胞进行了比较。随后,我们开发了一种使用实时 PCR 与特定探针的简单技术,以检查在口腔上皮样本中检测 OSCC 的能力,其中包括 103 个口腔冲洗液和 82 个口腔拭子样本。

结果

基于微切割组织的比较,NID2 的 cg22881914 被证明在大多数 OSCC 细胞中是甲基化的,但在正常口腔上皮细胞中是未甲基化的。此外,基于甲基化 NID2 的定量 PCR 方法已证明,该标志物有助于区分 OSCC 患者与正常口腔上皮细胞、吸烟者和口腔扁平苔藓患者,所用的样本是非侵入性的口腔冲洗液和拭子。

结论

OSCC 中 NID2 的 cg22881914 处的特异性甲基化可用作检测 OSCC 的重要潜在标志物。因此,需要进一步的研究来验证这个标志物的实用性,研究样本量还需要更大。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a69a/7071563/06f818cb362c/12885_2020_6692_Fig1_HTML.jpg

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