多重连接依赖探针扩增技术（MS-MLPA）在原发性喉鳞状细胞癌DNA甲基化谱分析中的应用
Utility of MS-MLPA in DNA methylation profiling in primary laryngeal squamous cell carcinoma.
作者信息
López Fernando, Sampedro Teresa, Llorente José L, Domínguez Francisco, Hermsen Mario, Suárez Carlos, Alvarez-Marcos César
机构信息
Department of Otorhinolaryngology and Instituto Universitario de Oncología del Principado de Asturias, Hospital Universitario Central de Asturias, Oviedo, Asturias, Spain.
Department of Medical Oncology, Hospital San Agustín, Avilés, Asturias, Spain.
出版信息
Oral Oncol. 2014 Apr;50(4):291-7. doi: 10.1016/j.oraloncology.2014.01.003. Epub 2014 Jan 18.
OBJECTIVES
Methylation-specific multiplex ligation-dependent probe amplification (MS-MLPA) assay is a method that has rarely been exploited in DNA methylation profiling of laryngeal squamous cell carcinoma (LSCC).
MATERIAL AND METHODS
Methylation of the gene was investigated by MS-MLPA in a well-characterized series of 53 LSCC and 30 samples of healthy mucosa. Aberrant promoter hypermethylation was confirmed using bisulfite pyrosequencing, and methylation-specific.
RESULTS
Promoter hypermethylation was observed in 36 of the 53 patients (68%). CDKN2B (28%), APC (17%), RARβ (15%), DAPK1 (11%) and CHFR (11%) were most frequently hypermethylated. Aberrant methylation of CHFR was mainly a late-stage event. Methylation-specific polymerase chain reaction and bisulfite pyrosequencing confirmed aberrant methylation for CDKN2B, APC and DAPK1.
CONCLUSION
Promoter methylation profiling of LSCC using MS-MLPA identified CDKN2B, DAPK1, RARβ, APC, and CHFR as frequent epigenetic events. The clinical implications of these genes as biomarkers are highly relevant as attractive targets for cancer therapy, given the reversible nature of epigenetic gene silencing.
目的
甲基化特异性多重连接依赖性探针扩增（MS-MLPA）检测是一种在喉鳞状细胞癌（LSCC）DNA甲基化谱分析中很少被应用的方法。
材料与方法
采用MS-MLPA对53例特征明确的LSCC样本和30例健康黏膜样本进行基因甲基化研究。使用亚硫酸氢盐焦磷酸测序法确认异常的启动子高甲基化，并进行甲基化特异性分析。
结果
53例患者中有36例（68%）观察到启动子高甲基化。CDKN2B（28%）、APC（17%）、RARβ（15%）、DAPK1（11%）和CHFR（11%）是最常发生高甲基化的基因。CHFR的异常甲基化主要是晚期事件。甲基化特异性聚合酶链反应和亚硫酸氢盐焦磷酸测序证实了CDKN2B、APC和DAPK1的异常甲基化。
结论
使用MS-MLPA对LSCC进行启动子甲基化谱分析发现，CDKN2B、DAPK1、RARβ、APC和CHFR是常见的表观遗传事件。鉴于表观遗传基因沉默的可逆性，这些基因作为生物标志物的临床意义与作为癌症治疗的有吸引力靶点高度相关。
Zotero 插件