Bosutinib induced cell differentiation and apoptosis in acute myeloid leukemia cell lines / 国际药学研究杂志

ABSTRACT

Objective To explore the effect of bosutinib on cell differentiation and apoptosis of human acute myeloid leukemia (AML) cell lines including HL-60, THP-1, and U937, and to clarify the related mechanisms. Methods MTT Assay was used to assess the proliferation of HL-60, THP-1, and U937 cells treated with various concentrations of bosutinib (0-20 μmol/L) for 48 h. Cell surface marker CD11b expression was detected by flow cytometry in HL-60, THP-1, and U937 cells treated with bosutinib (0-5 μmol/L) for 72 h. Apoptosis was evaluated by Annexin V-FITC/PI double stainning in the following two sets of experiments 1 HL- 60, THP-1 and U937 cells were treated with various concentrations of bosutinib (0-10 μmol/L). 2 U937 cells were pretreated with caspases inhibitor benzyloxy-carbonyl-Val-Ala-Asp-fluoromethylketone (Z-VAD-FMK) for 1 h and then exposed to bosutinib for 48 h. Western blot was used to examine the protein expression of differentiation related transcription factors C/EBPβ, P21, and c-Myc, and apoptosis related protein Mcl- 1, Bax, and Caspase 3 in HL- 60 and U937 cells treated with bosutinib for 24 h. Results Bosutinib dose- dependently inhibited AML cell growth. Treatment with various concentrations of bosutinib for 72 h significantly increased CD11b expression and apoptotic rate in AML cells as compared to blank control (P< 0.05 or P < 0.01). Bosutinib effectively upregulated protein expression levels of C/EBPβ and P21 in HL-60 cells, and induced down-regulation of Mcl-1 with up-regulation of Bax protein expression, and activated Caspase 3 in U937 ells. Pretreatment of U937 cells with Z-VAD-FMK significantly inhibited apoptosis induced by bosutinib. Conclusion Bosutinib effectively inhibits cell proliferation and induces cell differentiation with apoptosis in AML cells. It could be a potent therapeutic agent against AML.