口腔癌发生过程中N6-甲基腺苷（m6A）-RNA甲基化水平升高。

Elevated N6-Methyladenosine (m6A)-RNA-Methylation During Oral Carcinogenesis.

作者信息

Qin Zhiming, Chi Yanting, Wang Xinpei, Yan Jing, Zhang Xinning, Li Binbin

机构信息

Department of Oral Pathology, Peking University School and Hospital of Stomatology & National Center for Stomatology & National Clinical Research Center for Oral Diseases & National Engineering Research Center of Oral Biomaterials and Digital Medical Devices, Beijing, China.

Research Unit of Precision Pathologic Diagnosis in Tumors of the Oral and Maxillofacial Regions, Chinese Academy of Medical Sciences, Beijing, China.

出版信息

J Oral Pathol Med. 2025 Sep 22. doi: 10.1111/jop.70066.

DOI:10.1111/jop.70066

PMID:40984650

Abstract

BACKGROUND/PURPOSE: Oral epithelial dysplasia (OED) and oral squamous cell carcinoma (OSCC) encompass a series of molecular events in the malignant transformation process, ranging from simple epithelial hyperplasia to mild, moderate, and severe dysplasia. N6-methyladenosine (m6A)-RNA methylation participates in the regulation of the tumorigenesis of various malignant tumors, yet the roles played by m6A-RNA methylation in OED and OSCC remain unclear. Therefore, this study focused on investigating OED and OSCC from an epigenetic perspective, aiming to elucidate the underlying molecular mechanisms of malignant transformation.

MATERIALS AND METHODS

Laser microdissection was performed on OED and OSCC samples. Methylated RNA immunoprecipitation sequencing (MeRIP-Seq) and RNA sequencing (RNA-seq) were applied to establish comprehensive profiles of m6A methylation modifications and gene expression patterns and to identify differentially modified/expressed genes in OED and OSCC.

RESULTS

We presented the overall modification/expression profiles of m6A-RNA-methylation in OED and OSCC. Four hypermethylated genes and 11 hypomethylated genes were found in both OED and OSCC, together with the expression of 107 upregulated and 37 downregulated genes. The most common motifs GRAGRA (R = A/G) of the OED and OSCC methylation sites were mainly located in coding and stop codon regions. In the stable group, C4B, DNAH9, and NCALD all exhibited hypermethylated and upregulated, and the overall survival rate of patients with high expression of these genes was higher than that of patients with low-level expression.

CONCLUSION

Our study revealed that the level of m6A-RNA methylation in the epithelial tissues of OED and OSCC was higher than that in oral normal epithelium, suggesting that the methylation modification might be involved in the occurrence of OED and its progression to OSCC. Furthermore, hypermethylation and upregulated expression of C4B, DNAH9, and NCALD were associated with a favorable prognosis in these diseases.

摘要

背景/目的：口腔上皮发育异常（OED）和口腔鳞状细胞癌（OSCC）在恶性转化过程中包含一系列分子事件，范围从单纯上皮增生到轻度、中度和重度发育异常。N6-甲基腺苷（m6A）-RNA甲基化参与多种恶性肿瘤的肿瘤发生调控，但m6A-RNA甲基化在OED和OSCC中所起的作用仍不清楚。因此，本研究聚焦于从表观遗传学角度研究OED和OSCC，旨在阐明恶性转化的潜在分子机制。

材料与方法

对OED和OSCC样本进行激光显微切割。应用甲基化RNA免疫沉淀测序（MeRIP-Seq）和RNA测序（RNA-seq）建立m6A甲基化修饰和基因表达模式的综合图谱，并鉴定OED和OSCC中差异修饰/表达的基因。

结果

我们展示了OED和OSCC中m6A-RNA甲基化的整体修饰/表达图谱。在OED和OSCC中均发现4个高甲基化基因和11个低甲基化基因，同时有107个上调基因和37个下调基因表达。OED和OSCC甲基化位点最常见的基序GRAGRA（R = A/G）主要位于编码区和终止密码子区域。在稳定组中，C4B、DNAH9和NCALD均表现为高甲基化且上调，这些基因高表达患者的总生存率高于低表达患者。