Department of Molecular Cell Biology, Institute of Biochemistry, Life Sciences Center, Vilnius University, Vilnius, Lithuania.
Centre of Hematology, Oncology and Transfusion Medicine, Vilnius University Hospital Santaros Klinikos, Vilnius, Lithuania.
Mol Carcinog. 2019 Nov;58(11):2008-2016. doi: 10.1002/mc.23092. Epub 2019 Aug 6.
Treatment of acute myeloid leukemia (AML) is still a challenge because of common relapses or resistance to treatment. Therefore, the development of new therapeutic approaches is necessary. Various studies have shown that certain cancers, including some chemoresistant AML subsets, have upregulated oxidative phosphorylation. In this study, we aimed to assess treatment-resistant AML patients' cell modulation using oxidative phosphorylation inhibitors metformin and atovaquone alone and in various combinations with cytosine analog cytarabine and apoptosis inducer venetoclax. Metabolic activity analysis using Agilent Seahorse XF Extracellular Flux Analyzer revealed that peripheral blood mononuclear cells' metabolic state was different among treatment-resistant AML patients. We demonstrated that metformin decreased therapy-resistant-AML cell oxidative phosphorylation ex vivo, cotreatment with cytarabine and venetoclax slightly increased the effect. However, treatment with atovaquone did not have a marked effect in our experiment. Cell treatment had a slight effect on cell proliferation inhibition; combination of metformin, cytarabine, and venetoclax had the strongest effect. Moreover, a slightly higher effect on cell proliferation and cell cycle regulation was demonstrated in the cells with higher initial oxidative phosphorylation rate as demonstrated by gene expression analysis using reverse transcription quantitative polymerase chain reaction (RT-qPCR). Proteomic analysis by liquid chromatography-mass spectrometry demonstrated that chemoresistant AML cell treatment with metformin modulated metabolic pathways, while metformin combination with cytarabine and venetoclax boosted the effect. We suggest that oxidative phosphorylation inhibition is effective but not sufficient for chemoresistant AML treatment. Indeed, it causes anticancerous changes that might have an important additive role in combinatory treatment.
急性髓细胞白血病（AML）的治疗仍然是一个挑战，因为普遍存在复发或对治疗的耐药性。因此，有必要开发新的治疗方法。多项研究表明，某些癌症，包括一些化疗耐药的 AML 亚群，存在氧化磷酸化的上调。在这项研究中，我们旨在评估氧化磷酸化抑制剂二甲双胍和阿托伐醌单独以及与胞嘧啶类似物阿糖胞苷和凋亡诱导剂 venetoclax 联合治疗耐药性 AML 患者的细胞调节作用。使用安捷伦 Seahorse XF 细胞外通量分析仪进行代谢活性分析表明，耐药性 AML 患者的外周血单核细胞代谢状态不同。我们证明了二甲双胍可降低体外治疗耐药性 AML 细胞的氧化磷酸化，与阿糖胞苷和 venetoclax 联合治疗可略微增加其效果。然而，在我们的实验中，阿托伐醌的治疗没有明显的效果。细胞处理对细胞增殖抑制有轻微影响；二甲双胍、阿糖胞苷和 venetoclax 的联合治疗效果最强。此外，通过逆转录定量聚合酶链反应（RT-qPCR）进行基因表达分析表明，初始氧化磷酸化率较高的细胞的细胞增殖和细胞周期调节效果略高。通过液相色谱-质谱联用分析进行蛋白质组学分析表明，二甲双胍处理耐药性 AML 细胞可调节代谢途径，而二甲双胍与阿糖胞苷和 venetoclax 的联合治疗可增强这种作用。我们认为，氧化磷酸化抑制对耐药性 AML 的治疗是有效的，但还不够。事实上，它会引起抗癌变化，在联合治疗中可能具有重要的附加作用。
