Proliferation mechanism of acute myeloid leukemia cell line GDM-1 with CSF1R-Y571D mutation

Abstract

Objective To explore the proliferation mechanism of acute myeloid leukemia cell line GDM-1 by the method of protein kinase inhibition. Methods GDM-1 cells were divided into experimental group and control group according to different treatment. Experimental group was treated with 10 different protein kinase inhibitors, the control group was treated with DMSO, and the inhibition of cell proliferation was detected by CCK8 method. The phosphorylation level of key protein kinases in GDM-1 after treatment with linifanib, bosutinib, sorafenib were detected by Western blot.GDM-1 cells were treated with cytokine M-CSF alone or combined with protein kinase inhibitors, cell proliferation was detected by CCK8 method. Results Protein kinase inhibition analysis showed that dasatinib (IC50=0.01 μmol/L), bosutinib(IC50=0.07 μmol/L), linifanib (IC50=0.13 μmol/L) and sorafenib (IC50= 0.12 μmol/L) significantly inhibited the proliferation of GDM-1 cells. Western blot analysis showed that the phosphorylation level of Src and ERK in GDM-1 was decreased (P＜0.01) when treated with linifaninb. The proliferation of GDM-1 was 11.3 times as much as that of control group when stimulated with M-CSF. The proliferation of GDM-1 was dramatically inhibited when incubated with the combination of M-CSF and bosutinib (P＜0.001), but it was not significantly inhibited when incubated with M-CSF plus HG6-64-1. Conclusions CSF1R-Y571D mutation of GDM-1 cells increases the activity of CSF1R, and promotes cell proliferation by activating Src and MAPK signaling pathways.

Keywords

• protein kinase inhibitor|gdm-1|colony stimulating factor-1 receptor|linifanib|bosutinib