跨膜谷氨酰胺通量的竞争性拮抗剂V-9302对急性髓系白血病细胞系HL-60、KG-1凋亡的影响

Effect of Competitive Antagonist of Transmembrane Glutamine Flux V-9302 on Apoptosis of Acute Myeloid Leukemia Cell Lines HL-60 and KG-1

目的:研究跨膜谷氨酰胺通量的拮抗剂V-9302对急性髓系白血病细胞增殖、凋亡的影响。方法:取对数生长期的HL-60、KG-1细胞,给予不同浓度V-9302处理,CCK-8检测细胞增殖情况,采用Annexin V-FITC/PI双染色流式细胞术检测细胞的凋亡率,采用RT-qPCR、Western blot检测凋亡相关基因BAX、BCL-2、Caspase3的表达。结果:V-9302可以显著抑制HL-60、KG-1细胞的生长(P <0.05),10和20μmol/L的V-9302分别作用于HL-60、KG-1细胞系后,能显著促进细胞的凋亡(P <0.05);凋亡相关基因检测结果显示,10和20μmol/L的V-9302分别作用于HL-60、KG-1细胞后,促凋亡蛋白基因BAX、Caspase3表达水平显著高于对照组(P <0.05),而抗凋亡蛋白基因BCL-2的表达水平明显低于对照组(P <0.05),Western blot结果与RT-qPCR结果大致一致。结论:跨膜谷氨酰胺通量的竞争性拮抗剂V-9302能明显促进急性髓系白血病细胞系HL-60、KG-1的凋亡。

Objective:To investigate the effect of V-9302(an antagonist of transmembrane glutamine flux) on the proliferation and apoptosis of acute myeloid leukemia cells HL-60 and KG-1.Methods:HL-60 and KG-1 cells at logarithmic phase were treated by different concentrations of V-9302.CCK-8 assay was used to detect the proliferation of the cells.Annex in V-FITC/PI double staining flow cytometry was used to detect the apoptosis of HL-60 and KG-1 cells.The expressions of BAX,BCL-2 and Caspase3 were detected by RT-qPCR and Western blot.Results:V-9302 could significantly inhibit the growth of HL-60 and KG-1 cells.The concentration of V-9302 at 10,20 μmol/L could significantly promote the apoptosis of HL-60 and KG-1 cells(P <0.05).The results of apoptosis related gene detection showed that when V-9302 was applied to HL-60 and KG-1 cell lines at 10 and 20 μmol/L,the expression levels of Proapoptotic protein genes BAX and Caspase3 in HL-60 and KG-1 were significantly higher than those in control group(P <0.05),while the expression level of anti-apoptotic protein gene BCL-2 was significantly lower than that in the control group(P <0.05).The results of Western blot were basically consistent with that of RT-qPCR.Conclusion:Competitive antagonist of transmembrane glutamine flux V-9302 can significantly promote the apoptosis of acute myeloid leukemia cell lines HL-60 and KG-1.