Utility of MS-MLPA in DNA methylation profiling in primary laryngeal squamous cell carcinoma.
OBJECTIVE
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).


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.


CONCLUSIONS
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.
Department of Otorhinolaryngology and Instituto Universitario de Oncología del Principado de Asturias, Hospital Universitario Central de Asturias, Oviedo, Asturias, Spain. Electronic address: flopez_1981@yahoo.es.
Fernando
F
López
Department of Medical Oncology, Hospital San Agustín, Avilés, Asturias, Spain.
Teresa
T
Sampedro
Department of Otorhinolaryngology and Instituto Universitario de Oncología del Principado de Asturias, Hospital Universitario Central de Asturias, Oviedo, Asturias, Spain.
José L
JL
Llorente
Department of Pathology, Hospital de Cabueñes, Gijón, Asturias, Spain.
Francisco
F
Domínguez
Department of Otorhinolaryngology and Instituto Universitario de Oncología del Principado de Asturias, Hospital Universitario Central de Asturias, Oviedo, Asturias, Spain.
Mario
M
Hermsen
Department of Otorhinolaryngology and Instituto Universitario de Oncología del Principado de Asturias, Hospital Universitario Central de Asturias, Oviedo, Asturias, Spain.
Carlos
C
Suárez
Department of Otorhinolaryngology and Instituto Universitario de Oncología del Principado de Asturias, Hospital Universitario Central de Asturias, Oviedo, Asturias, Spain.
César
C
Alvarez-Marcos
D001483:Base Sequence; D002294:Carcinoma, Squamous Cell / Q000235:genetics*; D019175:DNA Methylation; D017931:DNA Primers; D006801:Humans; D007822:Laryngeal Neoplasms / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic
D017931:DNA Primers
CDKN2B; DNA methylation; Head and neck squamous cell carcinoma; Larynx squamous cell carcinoma; MS-MLPA