阻断PI3K/AKT/mTOR信号对白血病HL-60细胞的影响

Effect of Blocking PI3K/Akt/mTOR Signaling Transduction on Leukemia HL-60 Cells

目的探讨阻断PI3K/AKT/mTOR信号对白血病细胞的影响。方法选取急性粒细胞白血病细胞HL-60,随机分为空白对照组、低浓度组、中浓度组和高浓度组,其中空白对照组给予生理盐水,低、中、高浓度组分别给予10、25和50 mol/L浓度的PI3K/AKT/mTOR信号阻断剂GDC-0349处理,采用CCK-8法检测各组细胞增殖情况,流式细胞仪检测各组细胞凋亡情况,蛋白质印迹检测PI3K/AKT/mTOR信号、凋亡相关蛋白表达。结果高浓度组细胞培养24、48和72 h时吸光度值(A)明显低于低浓度组和中浓度组(P<0.05);低浓度组、中浓度组和高浓度组细胞凋亡率明显高于空白对照组(P<0.05),其中高浓度组细胞凋亡率明显低于低浓度组和中浓度组(P<0.05);高浓度组P110、AKT和mTOR蛋白相对表达量明显低于空白对照组、低浓度组和中浓度组(P<0.05);低浓度组、中浓度组和高浓度组Bcl2和Caspase3蛋白相对表达量明显低于空白对照组(P<0.05),高浓度组Bcl2和Caspase3蛋白相对表达量明显低于空白对照组、低浓度组和中浓度组(P<0.05)。结论阻断PI3K/AKT/mTOR信号可抑制白血病HL-60细胞增殖,促进细胞凋亡。

Objective To investigate the effect of blocking PI3K/Akt/mTOR signaling on leukemia cells.Methods Acute myeloid leukemia cell line HL-60 was selected,cells were randomly divided into 4 groups:blank control group,low concentration group,medium concentration group and high concentration group.Cells in the blank control group was given normal saline,cells in the low concentration group,medium concentration group and high concentration group were trea-ted with 10,25 and 50 mol/L PI3K/Akt/mTOR signaling blocker GDC-0349.CCK-8 method was used to detect the cell proliferation.Cell apoptosis was detected by flow cytometry,and the expression level of PI3K/Akt/mTOR and apoptosis related proteins were detected by Western blotting.Results Compared with those in the low and medium concentration groups,the absorbance ratios(A values)of cells in the high concentration group at 24,48 and 72 h time-points were significantly lower(P<0.05).The apoptosis rates of cells in the low,medium and high concentration groups were significantly higher than that of cells in the blank control group(P<0.05),and the apoptosis rate of cells in the high concentration group was significantly lower than those of cells in the low and medium concentration groups(P<0.05).The relative expression levels of P110,Akt and mTOR in the high concentration group were significantly lower than those in the blank control group,low and medium concentration groups(P<0.05).The relative expression levels of Bcl2 and Caspase3 proteins in the low,medium and high concentration groups were significantly lower than that in the blank control group(P<0.05).The relative expression levels of Bcl2 and Caspase3 in the high concentration group were significantly lower than those in other groups(P<0.05).Conclusion Blocking PI3K/Akt/mTOR signaling can inhibit the proliferation and promote the apoptosis of HL-60 cells.