采用甲基化敏感高分辨率熔解（MS-HRM）进行 DNA 甲基化分析，揭示了用于胶质瘤特征的基因panel。

DNA methylation analysis with methylation-sensitive high-resolution melting (MS-HRM) reveals gene panel for glioma characteristics.

机构信息

Department of Pharmaceutical Biochemistry, Poznan University of Medical Sciences, Poznań, Poland.

Department of Pediatric Gastroenterology and Metabolic Diseases, Poznan University of Medical Sciences, Poznań, Poland.

出版信息

CNS Neurosci Ther. 2020 Dec;26(12):1303-1314. doi: 10.1111/cns.13443. Epub 2020 Aug 11.

INTRODUCTION

Local DNA hypermethylation is a potential source of cancer biomarkers. While the evaluation of single gene methylation has limited value, their selected panel may provide better information.

AIMS

This study aimed to analyze the promoter methylation level in a 7-gene panel in brain tumors and verifies the usefulness of methylation-sensitive high-resolution melting (MS-HRM) for this purpose.

METHODS

Forty-six glioma samples and one non-neoplastic brain sample were analyzed by MS-HRM in terms of SFRP1, SFRP2, RUNX3, CBLN4, INA, MGMT, and RASSF1A promoter methylation. The results were correlated with patients' clinicopathological features.

RESULTS

DNA methylation level of all analyzed genes was significantly higher in brain tumor samples as compared to non-neoplastic brain and commercial, unmethylated DNA control. RASSF1A was the most frequently methylated gene, with statistically significant differences depending on the tumor WHO grade. Higher MGMT methylation levels were observed in females, whereas the levels of SFRP1 and INA promoter methylation significantly increased with patients' age. A positive correlation of promoter methylation levels was observed between pairs of genes, for example, CBLN4 and INA or MGMT and RASSF1A.

CONCLUSIONS

Our 7-gene panel of promoter methylation can be helpful in brain tumor diagnosis or characterization, and MS-HRM is a suitable method for its analysis.

简介

局部 DNA 高甲基化是癌症生物标志物的潜在来源。虽然单个基因甲基化的评估价值有限，但它们的选择组合可能提供更好的信息。

目的

本研究旨在分析脑肿瘤中 7 个基因启动子甲基化水平，并验证甲基化敏感高分辨率熔解（MS-HRM）在这方面的有效性。

方法

采用 MS-HRM 分析 46 例胶质瘤样本和 1 例非肿瘤性脑组织样本中 SFRP1、SFRP2、RUNX3、CBLN4、INA、MGMT 和 RASSF1A 启动子甲基化水平，并将结果与患者的临床病理特征相关联。

结果

与非肿瘤性脑组织和商用未甲基化 DNA 对照相比，所有分析基因的 DNA 甲基化水平在脑肿瘤样本中显著升高。RASSF1A 是最常发生甲基化的基因，其甲基化水平与肿瘤的 WHO 分级有统计学差异。MGMT 甲基化水平在女性中较高，而 SFRP1 和 INA 启动子甲基化水平随患者年龄的增加而显著增加。基因对之间的启动子甲基化水平呈正相关，例如 CBLN4 和 INA 或 MGMT 和 RASSF1A。