Department of Hematology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China.
Curr Med Sci. 2024 Aug;44(4):799-808. doi: 10.1007/s11596-024-2913-z. Epub 2024 Aug 3.
The metabolic reprogramming of acute myeloid leukemia (AML) cells is a compensatory adaptation to meet energy requirements for rapid proliferation. This study aimed to examine the synergistic effects of glutamine deprivation and metformin exposure on AML cells.
SKM-1 cells (an AML cell line) were subjected to glutamine deprivation and/or treatment with metformin or bis-2-(5-phenylacetamido-1,2,4-thiadiazol-2-yl) ethyl sulfide (BPTES, a glutaminase inhibitor) or cytarabine. Cell viability was detected by Cell Counting Kit-8 (CCK-8) assay, and cell apoptosis and reactive oxygen species (ROS) by flow cytometry. Western blotting was conducted to examine the levels of apoptotic proteins, including cleaved caspase-3 and poly(ADP-ribose) polymerase (PARP). Moreover, the human long noncoding RNA (lncRNA) microarray was used to analyze gene expression after glutamine deprivation, and results were confirmed with quantitative RT-PCR (qRT-PCR). The expression of metallothionein 2A (MT2A) was suppressed using siRNA. Cell growth and apoptosis were further detected by CCK-8 assay and flow cytometry, respectively, in cells with MT2A knockdown.
Glutamine deprivation or treatment with BPTES inhibited cell growth and induced apoptosis in SKM-1 cells. The lncRNA microarray result showed that the expression of MT family genes was significantly upregulated after glutamine deprivation. MT2A knockdown increased apoptosis, while proliferation was not affected in SKM-1 cells. In addition, metformin inhibited cell growth and induced apoptosis in SKM-1 cells. Both glutamine deprivation and metformin enhanced the sensitivity of SKM-1 cells to cytarabine. Furthermore, the combination of glutamine deprivation with metformin exhibited synergistic antileukemia effects on SKM-1 cells.
Targeting glutamine metabolism in combination with metformin is a promising new therapeutic strategy for AML.
急性髓系白血病（AML）细胞的代谢重编程是一种代偿性适应，以满足快速增殖所需的能量需求。本研究旨在研究谷氨酰胺剥夺和二甲双胍暴露对 AML 细胞的协同作用。
将 SKM-1 细胞（一种 AML 细胞系）进行谷氨酰胺剥夺和/或用二甲双胍或双-（5-苯乙酰氨基-1,2,4-噻二唑-2-基）乙硫醚（BPTES，谷氨酰胺酶抑制剂）或阿糖胞苷处理。通过细胞计数试剂盒-8（CCK-8）测定法检测细胞活力，通过流式细胞术检测细胞凋亡和活性氧（ROS）。通过 Western blot 检测凋亡蛋白（包括裂解的 caspase-3 和多聚（ADP-核糖）聚合酶（PARP））的水平。此外，使用人类长非编码 RNA（lncRNA）微阵列分析谷氨酰胺剥夺后的基因表达，并用定量 RT-PCR（qRT-PCR）进行验证。用 siRNA 抑制金属硫蛋白 2A（MT2A）的表达。在 MT2A 敲低的细胞中，通过 CCK-8 测定法和流式细胞术分别进一步检测细胞生长和凋亡。
谷氨酰胺剥夺或用 BPTES 处理抑制 SKM-1 细胞的生长并诱导其凋亡。lncRNA 微阵列结果表明，谷氨酰胺剥夺后 MT 家族基因的表达显著上调。MT2A 敲低增加了凋亡，而对 SKM-1 细胞的增殖没有影响。此外，二甲双胍抑制 SKM-1 细胞的生长并诱导其凋亡。谷氨酰胺剥夺和二甲双胍均增强了 SKM-1 细胞对阿糖胞苷的敏感性。此外，谷氨酰胺剥夺与二甲双胍联合对 SKM-1 细胞具有协同的抗白血病作用。
靶向谷氨酰胺代谢与二甲双胍联合是治疗 AML 的一种有前途的新治疗策略。
