title: Multistep pathogenesis of leukemia via the MLL-AF4 chimeric gene/Flt3 gene tyrosine kinase domain (TKD) mutation-related enhancement of S100A6 expression.
abstract: OBJECTIVE
  Concerning MLL-AF4 leukemogenesis, previous mouse models suggest that the tumorigenesis capacity of MLL-AF4 alone is insufficient for causing leukemia. Based on the finding that an Fms-like tyrosine kinase 3 (Flt3) gene mutation in the tyrosine kinase domain (TKD) was observed in approximately 15% of mixed lineage leukemia (MLL), we investigated synergistic leukemogenesis effects of the two genes in vitro.
  
  
  METHODS
  In a mouse interleukin-3 (IL-3)-dependent cell line, 32Dc, expression of MLL-AF4 and mutant Flt3 was induced using a lentiviral vector. We analyzed apoptosis induction in the absence of IL-3 and the granulocyte colony-stimulating factor-related induction of differentiation, gene expression profiling, and the mechanism involved in the synergistic effects of MLL-AF4 and Flt3-TKD.
  
  
  RESULTS
  Neither Flt3-expressing 32Dc (32Dc(Flt3-TKD)) nor MLL-AF4-expressing 32Dc (32Dc(MLL-AF4)) acquired IL-3-independent proliferative capacity in semisolid/liquid media. However, Flt3-TKD+MLL-AF4-expressing 32Dc (32Dc(Flt3-TKD+MLL-AF4)) acquired a non-IL-3-dependent proliferative capacity by inhibiting apoptosis in the two media. The 32Dc(Flt3-TKD) and 32Dc(MLL-AF4) cells differentiated into granulocytes in the presence of granulocyte colony-stimulating factor. However, in the 32Dc(Flt3-TKD+MLL-AF4) cells, there was no differentiation. Subsequently, we performed gene expression profiling. The enhancement of Hox genes expression was not identified. However, expression of S100A6 was synergistically enhanced in the presence of both MLL-AF4 and Flt3-TKD genes. Moreover, anti-S100A6 small interfering RNA downregulated leukemic proliferation.
  
  
  CONCLUSIONS
  We conclude that their synergistic enhancement of S100A6 expression plays an important role in MLL-AF4-associated leukemogenesis.
authors.affiliation: Division of Hematology, Department of Internal Medicine, Nippon Medical School, Bunkyo-Ku, Tokyo, Japan. y-hiroki@fd6.so-net.ne.jp
authors.forename: Hiroki
authors.initials: H
authors.lastname: Yamaguchi
authors.forename: Hideki
authors.initials: H
authors.lastname: Hanawa
authors.forename: Naoya
authors.initials: N
authors.lastname: Uchida
authors.forename: Mitsuharu
authors.initials: M
authors.lastname: Inamai
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Sawaguchi
authors.forename: Yoshio
authors.initials: Y
authors.lastname: Mitamura
authors.forename: Takashi
authors.initials: T
authors.lastname: Shimada
authors.forename: Kazuo
authors.initials: K
authors.lastname: Dan
authors.forename: Koiti
authors.initials: K
authors.lastname: Inokuchi
mesh_terms: D000818:Animals; D017209:Apoptosis; D020134:Catalytic Domain / Q000235:genetics; D018797:Cell Cycle Proteins / Q000032:analysis / Q000235:genetics*; D002460:Cell Line; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic; D016179:Granulocyte Colony-Stimulating Factor / Q000494:pharmacology; D007377:Interleukin-3 / Q000494:pharmacology; D007938:Leukemia / Q000209:etiology* / Q000235:genetics / Q000473:pathology; D051379:Mice; D009154:Mutation / Q000502:physiology*; D051788:Myeloid-Lymphoid Leukemia Protein / Q000235:genetics*; D015514:Oncogene Proteins, Fusion / Q000235:genetics*; D000075383:S100 Calcium Binding Protein A6; D009418:S100 Proteins / Q000032:analysis / Q000235:genetics*; D051941:fms-Like Tyrosine Kinase 3 / Q000235:genetics*
chemical_list: D018797:Cell Cycle Proteins; D007377:Interleukin-3; C113855:MLL-AF4 fusion protein, human; D015514:Oncogene Proteins, Fusion; D000075383:S100 Calcium Binding Protein A6; D009418:S100 Proteins; C489134:S100a6 protein, mouse; D016179:Granulocyte Colony-Stimulating Factor; D051788:Myeloid-Lymphoid Leukemia Protein; D051941:fms-Like Tyrosine Kinase 3