title: Sequential combination of decitabine and idarubicin synergistically enhances anti-leukemia effect followed by demethylating Wnt pathway inhibitor promoters and downregulating Wnt pathway nuclear target.
abstract: BACKGROUND
  The methylation inhibitor 5-Aza-2'-deoxycytidine (decitabine, DAC) has a great therapeutic value for acute myeloid leukemia (AML) and myelodysplastic syndromes (MDS). But decitabine monotherapy was associated with a relatively low rate of complete remission in AML and MDS. We aimed to investigate the effect of several anti-leukemia drugs in combination with decitabine on the proliferation of myeloid leukemia cells, to select the most efficient combination group and explore the associated mechanisms of these combination therapies.
  
  
  METHODS
  Cell proliferation was tested by MTT assay and CFU-GM assay. Cell apoptosis was evaluated by Annexin V and PI staining in cell culture, TUNEL assay and transmission electron microscopy in animal study. MicroPET was used to imaging the tumor in mouse model. Molecular studies were conducted using microarray expression analysis, which was used to explore associated pathways, and real-time quantitative reverse transcription-PCR, western blot and immunohistochemistry, used to assess regulation of Wnt/β-catenin pathway. Statistical significance among groups was determined by one-way ANOVA analysis followed by post hoc Bonferroni's multiple comparison test.
  
  
  RESULTS
  Among five anti-leukemia agents in combining with decitabine, the sequential combination of decitabine and idarubicin induced synergistic cell death in U937 cells, and this effect was verified in HEL, SKM-1 cells and AML cells isolated from AML patients. Importantly, tumor growth inhibition in this sequential combination was found to be higher than in single agent or controls in vivo. Moreover, sequential combination of the two agents induced apoptosis and depression of the Wnt/β-catenin pathway in both AML cell culture and animal studies.
  
  
  CONCLUSIONS
  The findings demonstrated that sequentially combination of decitabine and idarubicin had synergistic anti-leukemia effects. These effects were mainly attributed to demethylation of Wnt/β-catenin pathway inhibitors and downregulation of Wnt/β-catenin pathway nuclear targets.
authors.forename: Kongfei
authors.initials: K
authors.lastname: Li
authors.forename: Chao
authors.initials: C
authors.lastname: Hu
authors.forename: Chen
authors.initials: C
authors.lastname: Mei
authors.forename: Zhigang
authors.initials: Z
authors.lastname: Ren
authors.forename: Juan Carlos
authors.initials: JC
authors.lastname: Vera
authors.forename: Zhengping
authors.initials: Z
authors.lastname: Zhuang
authors.forename: Jie
authors.initials: J
authors.lastname: Jin
authors.affiliation: Department of Hematology, the First Affiliated Hospital of Zhejiang University, Hangzhou 310003, Zhejiang Province, People's Republic of China. hongyantong@aliyun.com.
authors.forename: Hongyan
authors.initials: H
authors.lastname: Tong
mesh_terms: D000818:Animals; D000971:Antineoplastic Combined Chemotherapy Protocols / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D001374:Azacitidine / Q000008:administration & dosage / Q000031:analogs & derivatives / Q000494:pharmacology; D002467:Cell Nucleus / Q000378:metabolism*; D049109:Cell Proliferation / Q000187:drug effects; D000077209:Decitabine; D015536:Down-Regulation; D004357:Drug Synergism; D006801:Humans; D015255:Idarubicin / Q000008:administration & dosage / Q000494:pharmacology; D020287:In Situ Nick-End Labeling; D007938:Leukemia / Q000473:pathology; D008745:Methylation; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D020298:U937 Cells; D051153:Wnt Proteins / Q000037:antagonists & inhibitors* / Q000378:metabolism
chemical_list: D051153:Wnt Proteins; D000077209:Decitabine; D001374:Azacitidine; D015255:Idarubicin