Low frequency of p16/CDKN2A methylation in sporadic melanoma: Comparative approaches to methylation analysis of primary tumors

Mark L. Gonzalgo, Christina M. Bender, Edward H. You, J. Michael Glendening, José F. Flores, Graeme J. Walker, Nicholas K. Hayward, Peter A. Jones, Jane W. Fountain

Research output: Contribution to journalArticlepeer-review

122 Scopus citations

Abstract

Methylation of the 5' CpG island of the p16 tumor suppressor gene represents one possible mechanism for inactivation of this cell cycle regulatory gene that is also a melanoma predisposition locus. We have investigated the potential contribution of somatic silencing of the p16 gene by DNA methylation in 30 cases of sporadic cutaneous melanoma. The methylation status of the 5' CpG island of p16 was initially determined by Southern analysis and then reevaluated (in a blinded manner) using methylation-specific PCR, methylation-sensitive single nucleotide primer extension, and bisulfite genomic sequencing. All methodologies yielded concordant results, and significant levels of methylation were observed in 3 of the 30 (10%) melanoma DNAs analyzed. Of the three tumors found to be methylated, two were also positive for LOH on 9p21 (where the p16 gene resides), implying that both p16 alleles were inactivated, one via deletion and the other via methylation-associated transcriptional silencing. The association between methylation and transcriptional silencing of p16 was also further supported by inducing p16 expression with a DNA demethylating agent (5-aza-2'-doexycytidine) in a melanoma cell line known to harbor a methylated p16 allele. Although methylation-associated gene silencing does not represent a common mechanism for p16 inactivation in sporadic melanoma, our findings provide support that PCR-based techniques, such as methylation-specific PCR and methylation-sensitive single nucleotide primer extension, can be reliably used for the accurate detection and quantitation of aberrant levels of DNA methylation in tumor specimens.

Original languageEnglish (US)
Pages (from-to)5336-5347
Number of pages12
JournalCancer Research
Volume57
Issue number23
StatePublished - Dec 1 1997
Externally publishedYes

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

Fingerprint

Dive into the research topics of 'Low frequency of p16/CDKN2A methylation in sporadic melanoma: Comparative approaches to methylation analysis of primary tumors'. Together they form a unique fingerprint.

Cite this