title: Lyn, a tyrosine kinase closely linked to the differentiation status of primary acute myeloid leukemia blasts, associates with negative regulation of all-trans retinoic acid (ATRA) and dihydroxyvitamin D3 (VD3)-induced HL-60 cells differentiation.
abstract: BACKGROUND
  Lyn, an import member of Src family kinases (SFKs), is supposed to be implicated in acute myeloid leukemia (AML) pathogenesis and development by participation in AML differentiation, yet the details still remain incompletely understood. The expression status of Lyn and its correlation with multiple clinical parameters including cell differentiation degree, different cytogenetic risk classification, and the activity of myeloperoxidase (MPO) were thus investigated. To address the mechanisms underlying the involvement of Lyn in differentiation induction, the effects of dasatinib, an inhibitor for SFKs including Lyn, on the alterations of all-trans retinoic acid (ATRA)- or dihydroxyvitamin D3 (VD3)-induced differentiation, and c-Myc protein expression were investigated.
  
  
  METHODS
  Primary AML blasts were obtained from 31 newly diagnosed AML patients with different French-American-British (FAB) subtypes. The expression of phosphorylated and total Lyn, c-Myc, and CD11b, CD11c and CD15 was analyzed by flow cytometry. The activation of Akt and Erk known to be involved in the regulation of c-Myc expression was investigated using western blotting.
  
  
  RESULTS
  Significant higher expression levels of total Lyn were observed in AML patients with favorable cytogenetics, higher MPO activity and FAB M2 subtype. A clear positive correlation between the expression levels of Lyn and differentiation status of primary AML blasts was observed. Dasatinib inhibited the expression of phosphorylated Lyn, and further enhanced the differentiation-inducing activity of ATRA and VD3 in HL-60 cells. Augmented downregulation of c-Myc protein expression was observed in the combination treatment with ATRA, VD3 and dasatinib compared to treatment with each reagent alone in HL-60 cells. The suppression of the activation of Akt and Erk was also observed concomitantly.
  
  
  CONCLUSIONS
  The expression level of total Lyn is closely linked to the differentiation status of AML blasts. The enhancement of differentiation-inducing activity of ATRA/VD3 by dasatinib suggested that Lyn was associated in the negative regulation of ATRA/VD3-induced HL-60 cells differentiation. The enhancement probably was attributed to the downregulation of c-Myc implicated with the suppression of the activation of Akt and Erk. These results provide novel insights into a possible combinational therapeutic approach by targeting Lyn for AML patients, and offer new possibilities for the combination therapy with VD3 and dasatinib.
authors.affiliation: Division of Hematology and Rheumatology, Department of Medicine, Nihon University School of Medicine, Itabashi Hospital, 30-1 Oyaguchi Kamicho, Itabashi-ku, Tokyo, Japan.
authors.forename: Noriyoshi
authors.initials: N
authors.lastname: Iriyama
authors.affiliation: Department of Applied Biochemistry, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, 1432-1 Horinouchi, Hachioji, Tokyo 192-0392 Japan.
authors.forename: Bo
authors.initials: B
authors.lastname: Yuan
authors.affiliation: Division of Hematology and Rheumatology, Department of Medicine, Nihon University School of Medicine, Itabashi Hospital, 30-1 Oyaguchi Kamicho, Itabashi-ku, Tokyo, Japan.
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Hatta
authors.affiliation: Department of Applied Biochemistry, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, 1432-1 Horinouchi, Hachioji, Tokyo 192-0392 Japan.
authors.forename: Norio
authors.initials: N
authors.lastname: Takagi
authors.affiliation: Division of Hematology and Rheumatology, Department of Medicine, Nihon University School of Medicine, Itabashi Hospital, 30-1 Oyaguchi Kamicho, Itabashi-ku, Tokyo, Japan.
authors.forename: Masami
authors.initials: M
authors.lastname: Takei
keywords: Acute myeloid leukemia; All-trans retinoic acid; Dihydroxyvitamin D3; Lyn; c-Myc; dasatinib