J Korean Assoc Oral Maxillofac Surg.  2005 Dec;31(6):468-473.

DNA methylation of TPEF gene in head and neck squamous cell carcinoma cell lines

• Chun SY
• Kim JO
• Hong SH
• Chung YK
• Jang HJ
• Shon YK
• Kim JW

Abstract

Head and neck squamous cell carcinoma (HNSCC) is the sixth most common malignancy worldwide. The molecular mechanisms involved in the development and progression of these carcinomas are not well known. Abnormalities of genomic methylation patterns have been attributed a role in carcinogenesis and local de novo methylation at tumor suppressor loci was held to be involved in silencing of tumor suppressor genes. Using Ms APPCR, we previously isolated a hypermethylated fragment corresponded to the 5'end of TPEF gene from primary liver and lung cancer cells. To confirm the inactivation of TPEF gene by hypermethylation in HNSCC, we investigated correlation between methylation pattern and expression of TPEF in 10 HNSCC cell lines. In methylation analysis such as combined-bisulfite restriction analysis(COBRA) and bisulfite sequencing, only RPMI 2650 showed none methylated pattern and another 9 cell lines showed dense methylation. The TPEF gene expression level analysis using RT-PCR showed that these 9 cell lines had not or significantly low expression levels of TPEF as compared with RPMI 2650. In addition, the increase of TPEF reexpression by 5-AzaC as demethylating agent in 9 cell lines also indicated that TPEF expression was regulated by hypermethylation. These results of this study demonstrate that epigenetic silencing of TPEF gene by aberrant methylation could play an important role in HNSCC carcinogenesis.

Keyword

Head and neck squamous cell carcinoma (HNSCC); DNA methylation; Combined-bisulfite restriction analysis(COBRA)

MeSH Terms

Carcinogenesis
Carcinoma, Squamous Cell*
Cell Line*
DNA Methylation*
DNA*
Epigenomics
Gene Expression
Genes, Tumor Suppressor
Head*
Liver
Lung Neoplasms
Methylation
Neck*
DNA