Effects of glycogen synthase kinase 3 inhibitors on the proliferation and apoptosis of chronic myelogenous leukemia K562 cells induced by imatinib / 白血病·淋巴瘤

ABSTRACT

Objective:To explore the effects of glycogen synthase kinase 3 (GSK3) inhibitors on the proliferation and apoptosis of chronic myelogenous leukemia (CML) K562 cells induced by imatinib.Methods:K562 cells were treated with 1 μmol/L imatinib combined with GSK3 inhibitor lithium chloride with different concentrations of 1.0, 2.0, 4.0 mmol/L, SB216763 with different concentrations of 0.5, 1.0, 5.0 μmol/L andTWS119 with different concentrations of 0.5, 1.0, 5.0 μmol/L, and then 1μmol/L imatinib was used as the control group. The proliferation activity of K562 cells was determined by using CCK8 assay. Flow cytometry was used to detect the cell apoptosis. The level changes of Wnt-β-catenin pathway related-protein were analyzed by using Western blot.Results:There were statistically significant differences of K562 cell survival rate between 1 μmol/L imatinib combined with different concentrations of SB216763, lithium chloride, TWS1193 groups and the control groups (all P < 0.01). The cell survival rate of 1 μmol/L imatinib + 1.0 μmol/L SB216763 group, 1 μmol/L imatinib + 5.0 μmol/L SB216763 group was (73.6±3.0)%, (77.0±3.6)%, which was higher than that of the control group [(68.0±2.8)%], and the difference was statistically significant (both P < 0.05). The cell survival rate of 1 μmol/L imatinib + 0.5 μmol/L SB216763 group was (70.0±2.2)%, and there was no statistical difference between 1 μmol/L imatinib + 0.5 μmol/L SB216763 group and the control group ( P > 0.05). The cell survival rate of 1 μmol/L imatinib + 2.0 mmol/L lithium chloride group and 1μmol/L imatinib + 4.0 mmol/L lithium chloride group was (75.5±3.6)%, (83.4±3.9)%, which was higher than that of the control group [(69.5±2.1)%], and the difference was statistically significant (both P < 0.05); there was no statistical difference in the cell survival rate of 1 μmol/L imatinib + 1.0 mmol/L lithium chloride group [(72.3±6.0)%] and the control group ( P > 0.05). The cell survival rate of 1 μmol/L imatinib combined with 0.5, 1.0, 5.0 μmol/L TWS119 was (70.0±1.1)%, (72.1±0.8)%, (73.8±0.7)%, respectively, which was higher than that of the control group [(67.9±7.5)%] (all P < 0.01). The cell apoptosis rate of 1 μmol/L imatinib + 5.0 μmol/L SB216763, 1 μmol/L imatinib + 4.0 mmol/L lithium chloride, 1 μmol/L imatinib + 5.0 μmol/L TWS119 was (18.16±3.59)%, (20.11±2.98)%, (16.27±2.36)%, respectively, which was lower than that of the control group [(28.26±2.20)%], and the difference was statistically significant (all P < 0.05). Compared with the imatinib group alone, there was no statistical difference in the protein expression levels of t-GSK3β, t-GSK3α of K562 cells treated with imatinib combined with GSK3 inhibitors, while the protein expression levels of p-GSK3β, p-GSK3α, β-catenin were increased. Conclusion:GSK3 inhibitors could reduce the effect of imatinib on the proliferation and apoptosis of CML K562 cells through regulating the related-protein level of Wnt-β-catenin pathway.