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

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.

Keywords: CDKN2B; DNA methylation; Head and neck squamous cell carcinoma; Larynx squamous cell carcinoma; MS-MLPA.

Publication types

  • Validation Study

MeSH terms

  • Base Sequence
  • Carcinoma, Squamous Cell / genetics*
  • DNA Methylation*
  • DNA Primers
  • Humans
  • Laryngeal Neoplasms / genetics*
  • Polymerase Chain Reaction
  • Promoter Regions, Genetic


  • DNA Primers