title: [Effect of Decitabine on DKK1 Gene Demethylation in Leukemia Cells].
abstract: OBJECTIVE
  To explore the effect of decitabine on Dickkopf-1 (DKK1) gene expression level and its downstream Wnt signaling pathway in acute myeloid leukemia (AML) cell line HL-60.
  
  
  METHODS
  Flow cytometry and DNA ladder analysis were performed to detect apoptosis in HL-60 cell treated with different concentration of decitabine. Methylation-specific polymerase chain reaction (MS-PCR) was used to examine the methylation status of DKK1 gene. The expressions of mRNA and protein were determined by qRT-PCR and Western blot, respectively.
  
  
  RESULTS
  Flow cytometric detection showed that after treating HL-60 cell line with decitabine of different concentrations for 48 h, the early apoptosis of HL-60 cells increased significantly as compared with control group (P < 0.05). DNA ladder analysis showed that the DNA ladder and demethylation of DKK1 gene appeared. RT-PCR and Western blot showed that the expressions of mRNA and protein increased. The protein expressions of β-catenin and C-MYC decreased.
  
  
  CONCLUSIONS
  The decitabine can promote the apoptosis of HL-60 cells throngh demethylation of DDK1 gene and inhibition of Wnt signalling pathway.
authors.affiliation: Department of Hematology, Guizhou Provincial People's Hospital, Guiyang 550002, Guizhou Province, China.
authors.forename: Feng-Zhi
authors.initials: FZ
authors.lastname: Liu
authors.affiliation: Department of Hematology, Affiliated Hospital of Guiyang Medical University, Guiyang 550002, Guizhou Province, China.
authors.forename: Ling
authors.initials: L
authors.lastname: He
authors.affiliation: Department of Hematology, Affiliated Hospital of Guiyang Medical University, Guiyang 550002, Guizhou Province, China.
authors.forename: Ji-Shi
authors.initials: JS
authors.lastname: Wang
authors.affiliation: Guangzhou Air Force Headquarters Hospital, Guangzhou 510071, Guangdong Province, China.
authors.forename: Song
authors.initials: S
authors.lastname: Zhang
authors.affiliation: Department of Hematology, Guizhou Provincial People's Hospital, Guiyang 550002, Guizhou Province, China. E-mail: zhuhongqian@126.com.
authors.forename: Hong-Qian
authors.initials: HQ
authors.lastname: Zhu
mesh_terms: D017209:Apoptosis; D001374:Azacitidine / Q000031:analogs & derivatives* / Q000494:pharmacology; D019175:DNA Methylation; D000077209:Decitabine; D015964:Gene Expression Regulation, Bacterial; D016259:Genes, myc; D018922:HL-60 Cells / Q000187:drug effects; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism*; D015470:Leukemia, Myeloid, Acute / Q000473:pathology*; D012333:RNA, Messenger; D060449:Wnt Signaling Pathway; D051176:beta Catenin / Q000378:metabolism
chemical_list: C495270:CTNNB1 protein, human; C501126:DKK1 protein, human; D036341:Intercellular Signaling Peptides and Proteins; D012333:RNA, Messenger; D051176:beta Catenin; D000077209:Decitabine; D001374:Azacitidine