title: [Effect of methylation inhibitor on EphB4 gene expression, proliferation and apoptosis in CEM cells].
abstract: OBJECTIVE
  To study the regulation of methylation inhibitor 5-aza-2'-deoxycytidine on transcription of EphB4 gene and effects on the proliferation and apoptosis of human acute lymphocyte leukemia cell line CEM.
  
  
  METHODS
  Bisulfite sequencing PCR was used to detect CpG island methylation density in EphB4 promoter. The expression of EphB4 mRNA and protein was determined by Q-PCR and Western blot. MTS assay and flow cytometry were used to detect the apoptosis of CEM cells after treatment with different concentrations of 5-aza-2'-deoxycytidine (1.0, 2.5 and 5 μmol/L).
  
  
  RESULTS
  Methylation of EphB4 gene promoter was detected in CEM cells (31.4%). The methylation level of EphB4 gene was down-regulated after treatment with various concentrations of 5-aza-2'-deoxycytidine. The EphB4 mRNA and protein expression in CEM cells increased after 5-aza-2'-deoxycytidine treatment. 5-Aza-2'-deoxycytidine significantly inhibited the cell growth in dose and time dependent manners. Early apoptosis rates of CEM cells increased from 4.1% to 24.8% 96 hrs after 5-aza-2'-deoxycytidine treatment. CEM cells in G1 phase decreased from 62.4% to 46.8%, cells in G2 phase increased from 2.1% to 16.2%, and CEM cells were arrested in G2 phase after treatment with 5 μmol/L 5-aza-2'-deoxycytidine for 96 hrs.
  
  
  CONCLUSIONS
  5-Aza-2'-deoxycytidine, an inhibitor of specific methylation transferase, can induce expression of the silent EphB4 gene in CEM cells, inhibit the proliferation of leukemia cells and induce cell apoptosis.
authors.affiliation: Department of Pediatrics, Second Affiliated Hospital of Jinan Medical College, Jinan University, Shenzhen, Guangdong 518020, China.
authors.forename: Yu-Hua
authors.initials: YH
authors.lastname: Li
authors.forename: Fei-Qiu
authors.initials: FQ
authors.lastname: Wen
authors.forename: Yi-Xin
authors.initials: YX
authors.lastname: Chen
authors.forename: Chang-Gang
authors.initials: CG
authors.lastname: Li
authors.forename: Zhao-Xia
authors.initials: ZX
authors.lastname: Zhang
authors.forename: Xiao-Wen
authors.initials: XW
authors.lastname: Chen
authors.forename: Bo
authors.initials: B
authors.lastname: Li
mesh_terms: D017209:Apoptosis / Q000187:drug effects*; D001374:Azacitidine / Q000031:analogs & derivatives* / Q000494:pharmacology; D002453:Cell Cycle / Q000187:drug effects; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D019175:DNA Methylation; D015254:DNA Modification Methylases / Q000037:antagonists & inhibitors*; D000077209:Decitabine; D006801:Humans; D054198:Precursor Cell Lymphoblastic Leukemia-Lymphoma / Q000188:drug therapy* / Q000473:pathology; D012333:RNA, Messenger / Q000032:analysis; D036224:Receptor, EphB4 / Q000235:genetics*
chemical_list: D012333:RNA, Messenger; D000077209:Decitabine; D015254:DNA Modification Methylases; D036224:Receptor, EphB4; D001374:Azacitidine