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. Annexin 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 Pro-apoptotic 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.
题目: 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.
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20 μmol/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20 μmol/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.