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Identification of candidate tumor suppressor genes inactivated by promoter methylation in melanoma

✍ Scribed by Vanessa F. Bonazzi; Darryl Irwin; Nicholas K. Hayward

Publisher
John Wiley and Sons
Year
2009
Tongue
English
Weight
400 KB
Volume
48
Category
Article
ISSN
1045-2257

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✦ Synopsis

Abstract

Tumor suppressor genes (TSGs) are sometimes inactivated by transcriptional silencing through promoter hypermethylation. To identify novel methylated TSGs in melanoma, we carried out global mRNA expression profiling on a panel of 12 melanoma cell lines treated with a combination of 5‐Aza‐2‐deoxycytidine (5AzadC) and an inhibitor of histone deacetylase, Trichostatin A. Reactivation of gene expression after drug treatment was assessed using Illumina whole‐genome microarrays. After qRT‐PCR confirmation, we followed up 8 genes (AKAP12, ARHGEF16, ARHGAP27, ENC1, PPP1R3C, PPP1R14C, RARRES1, and TP53INP1) by quantitative DNA methylation analysis using mass spectrometry of base‐specific cleaved amplification products in panels of melanoma cell lines and fresh tumors. PPP1R3C, ENC1, RARRES1, and TP53INP1, showed reduced mRNA expression in 35–59% of the melanoma cell lines compared to melanocytes and which was correlated with a high proportion of promoter methylation (>40–60%). The same genes also showed extensive promoter methylation in 6–25% of the tumor samples, thus confirming them as novel candidate TSGs in melanoma. © 2008 Wiley‐Liss, Inc.

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