Li Qian-Qin, Pan Shi-Yi, Chen Qian-Yi, Zhou Wei, Wang Shun-Qing
Department of Hematology, School of Medicine, South China University of Technology, Guangzhou 510006, Guangdong Province,China.
Guangzhou Medical University, Guangzhou 511436, Guangdong Province, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2021 Jun;29(3):685-689. doi: 10.19746/j.cnki.issn.1009-2137.2021.03.005.
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.
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.
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.
Competitive antagonist of transmembrane glutamine flux V-9302 can significantly promote the apoptosis of acute myeloid leukemia cell lines HL-60 and KG-1.
研究V - 9302(一种跨膜谷氨酰胺通量拮抗剂)对急性髓系白血病细胞HL - 60和KG - 1增殖及凋亡的影响。
用不同浓度的V - 9302处理对数期的HL - 60和KG - 1细胞。采用CCK - 8法检测细胞增殖情况。运用Annexin V - FITC/PI双染流式细胞术检测HL - 60和KG - 1细胞的凋亡情况。通过RT - qPCR和蛋白质免疫印迹法检测BAX、BCL - 2和Caspase3的表达。
V - 9302可显著抑制HL - 60和KG - 1细胞的生长。10、20 μmol/L的V - 9302浓度可显著促进HL - 60和KG - 1细胞凋亡(P<0.05)。凋亡相关基因检测结果显示,当10和20 μmol/L的V - 9302作用于HL - 60和KG - 1细胞系时,HL - 60和KG - 1中促凋亡蛋白基因BAX和Caspase3的表达水平显著高于对照组(P<0.05),而抗凋亡蛋白基因BCL - 2的表达水平显著低于对照组(P<0.05)。蛋白质免疫印迹法结果与RT - qPCR基本一致。
跨膜谷氨酰胺通量竞争性拮抗剂V - 9302可显著促进急性髓系白血病细胞系HL - 60和KG - 1的凋亡。