Synergistic Effects of Glutamine Deprivation and Metformin in Acute Myeloid Leukemia

Objective: 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.

Methods: 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.

Results: 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.

Conclusion: Targeting glutamine metabolism in combination with metformin is a promising new therapeutic strategy for AML.

acute myeloid leukemia; glutamine; metallothionein; metformin.