title: Non-promoter DNA hypermethylation of Zygote Arrest 1 (ZAR1) in neuroblastomas.
abstract: BACKGROUND
  The comprehensive methylation analysis of tumor-specific differently methylated regions in malignant melanomas and brain tumors has led to the identification of non-promoter hypermethylation of zygote arrest 1 (ZAR1). To search the non-promoter ZAR1 hypermethylation in neuroblastomas, we analyzed the levels of the methylation and transcript expression of ZAR1.
  
  
  METHODS
  The MassARRAY® EpiTYPER (Sequenom Inc., San Diego, CA, USA) system was optimized to determine the quantitative methylation levels of ZAR1 for 12 neuroblastoma cell lines, 23 neuroblastoma samples and four adrenal samples. ZAR1 expression levels were evaluated through a quantitative, real-time reverse transcription-polymerase chain reaction. The quantitative methylation levels of ZAR1 were subjected to correlation studies with the established markers of progressive disease and outcome.
  
  
  RESULTS
  Strikingly, the hypermethylation of ZAR1 regions and ZAR1 expression levels was observed in the neuroblastoma cell lines and neuroblastoma samples, compared to the adrenal samples. Somatic changes in ZAR1 methylation and ZAR1 expression were found in all three neuroblastoma patients. In the ZAR1 regions, poor-outcome tumors that were MYCN-amplified and/or Stage 3 or 4 and/or the age at diagnosis was≥18months, and/or showed an unfavorable histology were frequently hypermethylated.
  
  
  CONCLUSIONS
  Our results indicate that the hypermethylation of ZAR1 regions is extremely frequent in neuroblastomas and correlates with established markers of progressive disease and outcome.
authors.affiliation: Department of Pediatric Surgery, Nihon University School of Medicine, Tokyo, Japan. sugitou.kiminobu@nihon-u.ac.jp
authors.forename: Kiminobu
authors.initials: K
authors.lastname: Sugito
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Kawashima
authors.forename: Shinsuke
authors.initials: S
authors.lastname: Yoshizawa
authors.forename: Shota
authors.initials: S
authors.lastname: Uekusa
authors.forename: Reina
authors.initials: R
authors.lastname: Hoshi
authors.forename: Takeshi
authors.initials: T
authors.lastname: Furuya
authors.forename: Hide
authors.initials: H
authors.lastname: Kaneda
authors.forename: Toshifumi
authors.initials: T
authors.lastname: Hosoda
authors.forename: Noriyoshi
authors.initials: N
authors.lastname: Konuma
authors.forename: Takayuki
authors.initials: T
authors.lastname: Masuko
authors.forename: Kensuke
authors.initials: K
authors.lastname: Ohashi
authors.forename: Taro
authors.initials: T
authors.lastname: Ikeda
authors.forename: Tsugumichi
authors.initials: T
authors.lastname: Koshinaga
authors.forename: Ryouichi
authors.initials: R
authors.lastname: Tomita
authors.forename: Yui
authors.initials: Y
authors.lastname: Shinojima
authors.forename: Kyoko
authors.initials: K
authors.lastname: Fujiwara
authors.forename: Takao
authors.initials: T
authors.lastname: Watanabe
authors.forename: William A
authors.initials: WA
authors.lastname: Held
authors.forename: Hiroki
authors.initials: H
authors.lastname: Nagase
mesh_terms: D002648:Child; D002675:Child, Preschool; D019175:DNA Methylation; D018450:Disease Progression; D004527:Egg Proteins / Q000235:genetics*; D005260:Female; D006801:Humans; D007223:Infant; D008297:Male; D009367:Neoplasm Staging; D009447:Neuroblastoma / Q000235:genetics* / Q000473:pathology; D060888:Real-Time Polymerase Chain Reaction; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D018709:Statistics, Nonparametric; D015996:Survival Rate
chemical_list: D004527:Egg Proteins; C471631:Zar1 protein, human