Synergistic Effect of 3-Bromopyruvate in Combination with Rapamycin Impacted Neuroblastoma Metabolism by Inhibiting Autophagy.
Synergistic Effect of 3-Bromopyruvate in Combination with Rapamycin Impacted Neuroblastoma Metabolism by Inhibiting Autophagy.
复制标题
3-溴丙酮酸与雷帕霉素联用的协同作用通过抑制自噬影响神经母细胞瘤代谢
DOI：
10.2147/ott.s273108
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发表时间：
2020
影响因子：
4
通讯作者：
Zhuang Z
中科院分区：
文献类型：
--
作者：
Gan L;Ren Y;Lu J;Ma J;Shen X;Zhuang Z
细胞代谢的改变，如有氧糖酵解的增强，已被确定为癌细胞的一个突出标志。3-溴丙酮酸（3-BrPA）是已知的己糖激酶（HK）-II抑制剂，其可抑制癌细胞的能量代谢。雷帕霉素是一种新型的大环内酯类化合物，对丝氨酸/苏氨酸蛋白激酶mTOR具有抑制作用。为了了解3-BrPA对体外自噬活性的影响，我们使用不同的人神经母细胞瘤（NB）细胞系进行了一系列实验。材料与方法用3-BrPA和/或雷帕霉素处理人NB细胞株，CCK-8法检测细胞增殖活性。通过定量实时聚合酶链反应（QPCR）分析用3-BrPA和/或雷帕霉素处理的细胞的mRNA表达。Western Blotting（WB）法检测细胞蛋白表达。流式细胞仪检测3-BrPA和/或雷帕霉素对细胞周期和细胞凋亡的影响。同时，通过相关代谢分析试剂盒分析细胞葡萄糖吸收率、乳酸分泌率和ATP含量。结果3-BrPA可通过诱导细胞凋亡而抑制细胞生长，并呈剂量依赖性。3-BrPA联合雷帕霉素对NB细胞具有协同抑制作用，影响细胞凋亡、细胞周期和代谢途径。在与3-BrPA和雷帕霉素孵育的NB细胞中，上调的LC 3-II积累是有意识的。雷帕霉素单独抑制mTOR信号通路，而与3-BrPA组合可增强该现象并影响NB细胞的细胞代谢。结论3-BrPA联合雷帕霉素可通过抑制mTOR活性诱导NB细胞凋亡。总之，我们的研究表明，mTOR信号通路和糖酵解活性的双重抑制作用可能表明NB化学预防的有效治疗策略。
Background Alterations in the cell metabolism, such as enhanced aerobic glycolysis, have been identified as a prominent hallmark of cancer cells. 3-Bromopyruvate (3-BrPA) is a proverbial hexokinase (HK)-II inhibitor, which can inhibit cancer cell energy metabolism. Rapamycin is a new type macrocyclic lactone, which can inhibit the serine/threonine protein kinase mTOR. In order to comprehend the influence of 3-BrPA on autophagy activity in vitro, we conducted a series of experiments using different human neuroblastoma (NB) cell lines. Materials and Methods The human NB cell lines were exposed to 3-BrPA and/or rapamycin, and the proliferation activity of the cells was detected by Cell Counting Kit-8 (CCK-8) assay. The mRNA expression of the cells treated with 3-BrPA and/or rapamycin was analyzed by quantitative real-time polymerase chain reaction (QPCR) assay. The protein expression of the cells was analyzed by Western Blotting (WB) assay. The effects of 3-BrPA and/or rapamycin treatment on cell cycle and cell apoptosis were analyzed by flow cytometry assay. Meanwhile, the cellular glucose absorption rate, lactate secretion rate and ATP content were also analyzed through the relevant metabolic analysis kits. Results Our results showed that 3-BrPA can induce growth inhibition in a dose-dependent pattern by cell apoptosis. 3-BrPA combined with rapamycin played a synergistic suppression role in NB cells, affected the cell apoptosis, cell cycle and the metabolic pathway. Up-regulated LC3-II accumulation was conscious in NB cells incubated with 3-BrPA and rapamycin. Rapamycin individually discourages the mTOR signaling pathway, while combined with 3-BrPA can enhance this phenomenon and influence cell metabolism of the NB cells. Conclusion The results suggested that 3-BrPA combined with rapamycin could induce cell apoptosis in NB cells by inhibiting mTOR activity. In conclusion, our research proposed that the dual inhibitory effect of the mTOR signaling pathway and the glycolytic activity may indicate a valid therapeutic tactic for NB chemoprevention.