title: Even-skipped homeobox 1 is frequently hypermethylated in prostate cancer and predicts PSA recurrence.
abstract: BACKGROUND
  DNA methylation is an important epigenetic mechanism in prostate cancer (PCa) progression. Given the role of even-skipped homeobox 1 (EVX1) in the regulation of multiple genes during embryogenesis, we postulated that EVX1 methylation is altered in PCa progression.
  
  
  METHODS
  Bisulphite sequencing and quantitative MethyLight were used to assess methylation in human prostate epithelial cells, four PCa cell lines, liver, lung, spleen, kidney, 35 paired tumour and tumour-associated benign tissues, and 11 normal prostate tissues. Prostate cancer cell lines were treated with 5-azacytidine (AzaC) or trichostatin A (TSA), and expression of EVX1 transcript and variants was assessed by qPCR. Hypermethylation was compared with clinicopathological features in a validation set of 58 patients using microarray.
  
  
  RESULTS
  Even-skipped homeobox 1 hypermethylation was observed in all four PCa cell lines and 57% of tumours. High-grade tumours exhibited increased methylation compared with intermediate-grade tumours. Even-skipped homeobox 1 expression was induced in PCa cell lines after treatment with AzaC or TSA. In the validation set, 83% of tumours were hypermethylated and hypermethylation was associated with worse recurrence-free survival.
  
  
  CONCLUSIONS
  In this first evaluation of EVX1 methylation in human cancer, EVX1 is one of the most commonly hypermethylated genes observed in PCa and predicted treatment failure in moderate risk patients.
authors.affiliation: Department of Urology, University of Wisconsin School of Medicine and Public Health, 7037, Wisconsin Institutes for Medical Research, 1111 Highland Avenue, Madison, WI 53792, USA.
authors.forename: M
authors.initials: M
authors.lastname: Truong
authors.forename: B
authors.initials: B
authors.lastname: Yang
authors.forename: J
authors.initials: J
authors.lastname: Wagner
authors.forename: Y
authors.initials: Y
authors.lastname: Kobayashi
authors.forename: V
authors.initials: V
authors.lastname: Rajamanickam
authors.forename: J
authors.initials: J
authors.lastname: Brooks
authors.forename: D F
authors.initials: DF
authors.lastname: Jarrard
mesh_terms: D001374:Azacitidine / Q000494:pharmacology; D045744:Cell Line, Tumor; D019175:DNA Methylation; D018572:Disease-Free Survival; D004791:Enzyme Inhibitors / Q000494:pharmacology; D056572:Histone Deacetylase Inhibitors / Q000494:pharmacology; D018398:Homeodomain Proteins / Q000235:genetics*; D006801:Humans; D006877:Hydroxamic Acids / Q000494:pharmacology; D008297:Male; D011379:Prognosis; D011467:Prostate / Q000378:metabolism; D017430:Prostate-Specific Antigen / Q000378:metabolism; D011471:Prostatic Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D012008:Recurrence
chemical_list: D004791:Enzyme Inhibitors; D056572:Histone Deacetylase Inhibitors; D018398:Homeodomain Proteins; D006877:Hydroxamic Acids; C074437:EVX1 protein, human; C012589:trichostatin A; D017430:Prostate-Specific Antigen; D001374:Azacitidine