Metabolomics profiling of metformin-mediated metabolic reprogramming bypassing AMPK alpha
Metabolomics profiling of metformin-mediated metabolic reprogramming bypassing AMPK alpha
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二甲双胍介导的绕过 AMPK α 的代谢重编程的代谢组学分析
DOI：
10.1016/j.metabol.2018.11.010
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发表时间：
2019
影响因子：
9.8
通讯作者：
Piao Hai long
中科院分区：
文献类型：
--
作者：
Yan Min;Qi Huan;Xia Tian;Zhao Xinjie;Wang Wen;Wang Zhichao;Lu Chang;Ning Zhen;Chen Huan;Li Tongming;Tekcham Dinesh Singh;Liu Xiumei;Liu Jing;Chen Di;Liu Xiaolong;Xu Guowang;Piao Hai long
背景二甲双胍是治疗2型糖尿病的一线药物，作为一种潜在的抗癌药物已引起人们的极大兴趣。越来越多的证据表明，二甲双胍通过破坏代谢稳态和改变能量状态来拮抗糖尿病和肿瘤。然而，在这方面，方法采用气相色谱-质谱联用（GC/MS）和毛细管电泳-质谱联用（CE-MS）技术，系统研究二甲双胍对Prkaa 1野生型（WT）和基因敲除（KO）小鼠胚胎成纤维细胞（MEF）以及人细胞代谢途径的影响。结果二甲双胍诱导Prkaa 1 WT和KO MEF细胞代谢重编程，并降低细胞能量状态，表现为抑制三羧酸（TCA）循环、增加乳酸产量以及降低NAD+/NADH比值。此外，代谢通量分析还显示二甲双胍Ampkα独立地增加了从葡萄糖到乳酸的代谢通量，降低了从乙酰辅酶A到TCA循环以及从丙酮酸到苹果酸的代谢通量。此外，二甲双胍Ampkα依赖性上调MEF细胞中P-Acc，但Ampkα独立地抑制MEF细胞中P-mTor、P-S6、Lc 3、Atgl和P-Erk的水平。同样，我们证明了常用的AMPK激动剂5-氨基咪唑-4-甲酰胺核苷酸（AICAR）和胎牛血清（FBS）饥饿，作为一种常见的能量应激模型，都导致MEF细胞中Ampkα非依赖性代谢的改变。此外，二甲双胍的这些作用也在人肝细胞癌（HCC）细胞以及MCF 10A shControl和shPRKAA 1细胞中得到证实。重要的是，我们发现，二甲双胍可以明显抑制肝癌细胞的集落构象在Ampkα-独立的martens.ConclusionsOur数据突出了二甲双胍以及AICAR介导的代谢重编程的全面看法。这些观察结果表明，二甲双胍可以在很大程度上绕过Ampkα影响细胞代谢，并可能为其临床应用提供新的见解。
BackgroundMetformin is a first-line drug for treating type 2 diabetes and has gained considerable interest as a potential anticancer agent. Increasing evidence suggests that metformin antagonizes diabetes and tumors through disrupting metabolic homeostasis and altering energy state. However, whether AMP activated protein kinase (AMPK) contributes to such effects of metformin remains controversial.MethodsWe performed integrative metabolomics analyses to systematically examine the effects of metformin on metabolic pathways inPrkaa1wild type (WT) and knock-out (KO) mouse embryonic fibroblast (MEF) cells as well as human cells based on gas chromatography–mass spectrometry and capillary electrophoresis–mass spectrometry (CE–MS).ResultsMetformin treatment induced metabolic reprogramming and reduced the energy state of bothPrkaa1WT and KO MEF cells, as evidenced by suppressed tricarboxylic acid (TCA) cycle, elevated lactate production as well as decreased NAD+/NADH ratio. Additionally, metabolic flux analysis also showed that metformin Ampkα-independently increased metabolic flux from glucose to lactate and decreased metabolic flux from acetyl-CoA to TCA cycle as well as from pyruvate to malate. Moreover, metformin Ampkα-dependently upregulated P-Acc but Ampkα-independently inhibited the levels of P-mTor, P-S6, Lc3, Atgl and P-Erk in MEF cells. Similarly, we demonstrated that a commonly used AMPK agonist 5-Aminoimidazole-4-carboxamide ribonucleotide (AICAR) and fetal bovine serum (FBS) starvation, as a common model for energy stress, both led to Ampkα-independent metabolism alterations in MEF cells. Furthermore, these effects of metformin were also confirmed in human hepatocellular carcinoma (HCC) cells as well as in MCF10A shControl and shPRKAA1 cells. Importantly, we found that metformin could obviously inhibit colony conformation of HCC cells in an Ampkα-independent manner.ConclusionsOur data highlight a comprehensive view of metabolic reprogramming mediated by metformin as well as AICAR. These observations suggest that metformin could affect cellular metabolism largely bypassing Ampkα, and may provide a new insight for its clinical usage.