Department of Pediatrics, Renmin Hospital of Wuhan University, Wuhan, Hubei, China (mainland).
Hubei Medical Devices, Quality Supervision and Test Institute, Wuhan, Hubei, China (mainland).
Med Sci Monit. 2020 May 29;26:e920810. doi: 10.12659/MSM.920810.
BACKGROUND Nicotinamide can affect differentiation and proliferation of leukemia cells. This research aimed to explore the regulatory effect of nicotinamide on glycolysis metabolism of leukemia cells and to clarify the associated mechanisms. MATERIAL AND METHODS HL-60 cells were treated with nicotinamide and divided into 0.1, 1, and 10 μmol/l groups. HL-60 cells without any administration were assigned as negative control (CT group). Glucolytic activity was evaluated by detecting lactic acid production, and glucose level was measured using glucose consumption assay. Apoptosis of HL-60 was examined using flow cytometry assay, when cells were cultured for 24 h. Expressions of sirtuin 1 (SIRT1), peroxisome proliferator-activated receptor γ coactivator 1alpha (PGC-1alpha), and hypoxia-inducible factor-2alpha (HIF2alpha) were evaluated using a reverse transcription PCR assay and Western blotting assay, respectively. RESULTS Nicotinamide remarkably decreased lactic acid production and glucose levels in leukemia cells compared with that of the CT group (p<0.05). Nicotinamide significantly induced the apoptosis of HL-60 cells compared to that of the negative control group (p<0.05). Nicotinamide significantly inhibited the SIRT1/PGC-1alpha/HIF2alpha signaling pathway mRNAs compared to that of the CT group (p<0.05). Nicotinamide remarkably reduced mitochondrial regulatory factors SIRT1/PGC-1alpha expression compared to that in the CT group (p<0.05). Nicotinamide obviously downregulated HIF2alpha compared with that of the CT group (p<0.05). Moreover, all of the above nicotinamide-induced effects, including glycolytic activity, apoptosis, and expression of SIRT1/PGC-1alpha/HIF2alpha, were changed in a dose-dependent manner. CONCLUSIONS Nicotinamide can inhibit glycolysis of HL-60 cells by inhibiting the mitochondrial regulatory factor SIRT1/PGC-1alpha and suppressing transcription factor HIF2alpha.
烟酰胺可以影响白血病细胞的分化和增殖。本研究旨在探讨烟酰胺对白血病细胞糖酵解代谢的调节作用，并阐明其相关机制。
用烟酰胺处理 HL-60 细胞，并分为 0.1、1 和 10μmol/L 组。未进行任何处理的 HL-60 细胞被分配为阴性对照（CT 组）。通过检测乳酸产量评估糖酵解活性，通过葡萄糖消耗测定法测量葡萄糖水平。当细胞培养 24 小时时，使用流式细胞术检测 HL-60 细胞的凋亡。使用逆转录 PCR 法和 Western blot 法分别评估沉默信息调节因子 1（SIRT1）、过氧化物酶体增殖物激活受体γ共激活因子 1α（PGC-1α）和缺氧诱导因子-2α（HIF2α）的表达。
与 CT 组相比，烟酰胺显著降低白血病细胞的乳酸产量和葡萄糖水平（p<0.05）。与阴性对照组相比，烟酰胺显著诱导 HL-60 细胞凋亡（p<0.05）。与 CT 组相比，烟酰胺显著抑制 SIRT1/PGC-1α/HIF2α 信号通路的 mRNA（p<0.05）。与 CT 组相比，烟酰胺显著降低线粒体调节因子 SIRT1/PGC-1α 的表达（p<0.05）。与 CT 组相比，烟酰胺明显下调 HIF2α（p<0.05）。此外，烟酰胺诱导的所有上述作用，包括糖酵解活性、凋亡和 SIRT1/PGC-1α/HIF2α 的表达，均呈剂量依赖性变化。
烟酰胺通过抑制线粒体调节因子 SIRT1/PGC-1α 和抑制转录因子 HIF2α，抑制 HL-60 细胞的糖酵解。
