title: Valproate induces widespread epigenetic reprogramming which involves demethylation of specific genes.
abstract: Valproate (VPA)(1) has been used for decades in the treatment of epilepsy, and is also effective as a mood stabilizer and in migraine therapy. It has been shown that VPA is also a histone deacetylase (HDAC) inhibitor. We have previously shown that VPA could trigger active demethylation of ectopically methylated transiently transfected DNA in HEK 293 cells. We therefore tested whether VPA treatment could bring about stable changes in the epigenome by causing changes in the state of DNA methylation of genomic DNA. Using a microarray gene expression analysis we identified the genes whose expression is induced by VPA treatment in HEK 293 cells. We found that a subset of these genes could also be induced by the classical DNA methylation inhibitor 5-aza-2'-deoxy-cytidine (5-aza-CdR) suggesting that VPA can alter the state of expression of genes, which are stably suppressed by DNA methylation. We mapped the state of methylation of three of these genes, MELANOMA ANTIGEN B2 GENE (MAGEB2), METALLOPROTEINASE 2 (MMP2) and WIF1, which are involved in tumor growth and metastasis. A chromatin immunoprecipitation (ChIP) assay revealed that VPA treatment caused as expected a change in the state of acetylation of these genes. Our data supports the concept that chromatin acetylation and DNA methylation are found in a dynamic interrelation and that the consequences of HDAC inhibitors are not limited to changes in histone acetylation but that they also bring about a change in the state of modification of DNA. The implications of our results on the future therapeutic utilities of VPA in cancer will be discussed.
authors.affiliation: Department of Pharmacology and Therapeutics, McGill University, Montreal, Canada.
authors.forename: Snezana
authors.initials: S
authors.lastname: Milutinovic
authors.forename: Ana C
authors.initials: AC
authors.lastname: D'Alessio
authors.forename: Nancy
authors.initials: N
authors.lastname: Detich
authors.forename: Moshe
authors.initials: M
authors.lastname: Szyf
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics; D000951:Antigens, Neoplasm / Q000235:genetics; D002460:Cell Line; D002843:Chromatin / Q000235:genetics; D019175:DNA Methylation / Q000187:drug effects; D044127:Epigenesis, Genetic / Q000187:drug effects* / Q000235:genetics*; D056572:Histone Deacetylase Inhibitors; D006655:Histone Deacetylases / Q000235:genetics; D006801:Humans; D007668:Kidney; D020778:Matrix Metalloproteinase 2 / Q000235:genetics; D009363:Neoplasm Proteins / Q000235:genetics; D012097:Repressor Proteins / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014635:Valproic Acid / Q000494:pharmacology*
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D000951:Antigens, Neoplasm; D002843:Chromatin; D056572:Histone Deacetylase Inhibitors; C072012:MAGEB2 protein, human; D009363:Neoplasm Proteins; D012097:Repressor Proteins; C485569:WIF1 protein, human; D014635:Valproic Acid; D020778:Matrix Metalloproteinase 2; D006655:Histone Deacetylases