Department of Pediatrics, Qilu Hospital of Shandong University, Jinan, 250012, Shandong Province, China.
Laboratory of Cryomedicine, Qilu Hospital of Shandong University, Jinan, 250012, Shandong Province, China.
BMC Cancer. 2023 Feb 6;23(1):122. doi: 10.1186/s12885-023-10589-9.
Precursor B-cell acute lymphoblastic leukemia (pre-B ALL) is the most common hematological malignancy in children. Cellular metabolic reorganization is closely related to the progression and treatment of leukemia. We found that the level of 1,5-anhydroglucitol (1,5-AG), which is structurally similar to glucose, was elevated in children with pre-B ALL. However, the effect of 1,5-AG on pre-B ALL was unclear. Here, we aimed to reveal the roles and mechanisms of 1,5-AG in pre-B ALL progression.
The peripheral blood plasma level of children with initial diagnosis of pre-B ALL and that of healthy children was measured using untargeted metabolomic analysis. Cell Counting Kit-8 assay, RNA sequencing, siRNA transfection, real-time quantitative PCR, and western blot were performed using pre-B ALL cell lines Reh and HAL-01. Cell cycle, cell apoptosis, ROS levels, and the positivity rate of CD19 were assessed using flow cytometry. Oxygen consumption rates and extracellular acidification rate were measured using XFe24 Extracellular Flux Analyzer. The lactate and nicotinamide adenine dinucleotide phosphate levels were measured using kits. The effect of 1,5-AG on pre-B ALL progression was verified using the In Vivo Imaging System in a xenotransplantation leukemia model.
We confirmed that 1,5-AG promoted the proliferation, viability, and intracellular glycolysis of pre-B ALL cells. Mechanistically, 1,5-AG promotes glycolysis while inhibiting mitochondrial respiration by upregulating pyruvate dehydrogenase kinase 4 (PDK4). Furthermore, high levels of intracellular glycolysis promote pre-B ALL progression by activating the reactive oxygen species (ROS)-dependent mitogen-activated protein kinase/extracellular signal-regulated kinase (MAPK/ERK) pathway. Conversely, N-acetylcysteine or vitamin C, an antioxidant, effectively inhibited 1,5-AG-mediated progression of leukemia cells.
Our study reveals a previously undiscovered role of 1,5-AG in pre-B ALL, which contributes to an in-depth understanding of anaerobic glycolysis in the progression of pre-B ALL and provides new targets for the clinical treatment of pre-B ALL.
前 B 细胞急性淋巴细胞白血病（pre-B ALL）是儿童中最常见的血液系统恶性肿瘤。细胞代谢重排与白血病的进展和治疗密切相关。我们发现，结构上类似于葡萄糖的 1,5-脱水葡萄糖醇（1,5-AG）的水平在患有 pre-B ALL 的儿童中升高。然而，1,5-AG 对 pre-B ALL 的影响尚不清楚。在这里，我们旨在揭示 1,5-AG 在 pre-B ALL 进展中的作用和机制。
使用非靶向代谢组学分析测量初诊 pre-B ALL 患儿和健康儿童的外周血血浆水平。使用 pre-B ALL 细胞系 Reh 和 HAL-01 进行细胞计数试剂盒-8 测定、RNA 测序、siRNA 转染、实时定量 PCR 和 Western blot。使用流式细胞术评估细胞周期、细胞凋亡、ROS 水平和 CD19 的阳性率。使用 XFe24 细胞外通量分析仪测量耗氧率和细胞外酸化率。使用试剂盒测量乳酸和烟酰胺腺嘌呤二核苷酸磷酸水平。使用体内成像系统在异种移植白血病模型中验证 1,5-AG 对 pre-B ALL 进展的影响。
我们证实 1,5-AG 促进 pre-B ALL 细胞的增殖、活力和细胞内糖酵解。从机制上讲，1,5-AG 通过上调丙酮酸脱氢酶激酶 4（PDK4）促进糖酵解，同时抑制线粒体呼吸。此外，高水平的细胞内糖酵解通过激活活性氧（ROS）依赖性丝裂原活化蛋白激酶/细胞外信号调节激酶（MAPK/ERK）通路促进 pre-B ALL 的进展。相反，抗氧化剂 N-乙酰半胱氨酸或维生素 C 可有效抑制 1,5-AG 介导的白血病细胞进展。
本研究揭示了 1,5-AG 在 pre-B ALL 中的一个以前未被发现的作用，有助于深入了解无氧糖酵解在 pre-B ALL 进展中的作用，并为 pre-B ALL 的临床治疗提供了新的靶点。
