Synergy of demethylation and histone deacetylase inhibition in the re-expression of genes silenced in cancer

Nat Genet. 1999 Jan;21(1):103-7. doi: 10.1038/5047.

Abstract

Densely methylated DNA associates with transcriptionally repressive chromatin characterized by the presence of underacetylated histones. Recently, these two epigenetic processes have been dynamically linked. The methyl-CpG-binding protein MeCP2 appears to reside in a complex with histone deacetylase activity. MeCP2 can mediate formation of transcriptionally repressive chromatin on methylated promoter templates in vitro, and this process can be reversed by trichostatin A (TSA), a specific inhibitor of histone deacetylase. Little is known, however, about the relative roles of methylation and histone deacetylase activity in the stable inhibition of transcription on densely methylated endogenous promoters, such as those for silenced alleles of imprinted genes, genes on the female inactive X chromosome and tumour-suppressor genes inactivated in cancer cells. We show here that the hypermethylated genes MLH1, TIMP3 (TIMP3), CDKN2B (INK4B, p15) and CDKN2A (INK4, p16) cannot be transcriptionally reactivated with TSA alone in tumour cells in which we have shown that TSA alone can upregulate the expression of non-methylated genes. Following minimal demethylation and slight gene reactivation in the presence of low dose 5-aza-2'deoxycytidine (5Aza-dC), however, TSA treatment results in robust re-expression of each gene. TSA does not contribute to demethylation of the genes, and none of the treatments alter the chromatin structure associated with the hypermethylated promoters. Thus, although DNA methylation and histone deacetylation appear to act as synergistic layers for the silencing of genes in cancer, dense CpG island methylation is dominant for the stable maintenance of a silent state at these loci.

MeSH terms

  • Adaptor Proteins, Signal Transducing
  • Azacitidine / analogs & derivatives
  • Azacitidine / pharmacology
  • Carrier Proteins / genetics
  • Cell Cycle Proteins*
  • CpG Islands
  • Cyclin-Dependent Kinase Inhibitor p15
  • Cyclin-Dependent Kinase Inhibitor p16 / genetics
  • DNA Methylation*
  • DNA Modification Methylases / antagonists & inhibitors
  • Decitabine
  • Enzyme Inhibitors / pharmacology
  • Gene Expression Regulation, Neoplastic / drug effects*
  • Histone Deacetylase Inhibitors*
  • Humans
  • Hydroxamic Acids / pharmacology*
  • MutL Protein Homolog 1
  • Neoplasm Proteins / genetics
  • Neoplasms / genetics
  • Nuclear Proteins
  • Promoter Regions, Genetic
  • Tissue Inhibitor of Metalloproteinase-3 / genetics
  • Transcriptional Activation / drug effects
  • Tumor Cells, Cultured
  • Tumor Suppressor Proteins*

Substances

  • Adaptor Proteins, Signal Transducing
  • CDKN2B protein, human
  • Carrier Proteins
  • Cell Cycle Proteins
  • Cyclin-Dependent Kinase Inhibitor p15
  • Cyclin-Dependent Kinase Inhibitor p16
  • Enzyme Inhibitors
  • Histone Deacetylase Inhibitors
  • Hydroxamic Acids
  • MLH1 protein, human
  • Neoplasm Proteins
  • Nuclear Proteins
  • Tissue Inhibitor of Metalloproteinase-3
  • Tumor Suppressor Proteins
  • trichostatin A
  • Decitabine
  • DNA Modification Methylases
  • MutL Protein Homolog 1
  • Azacitidine