Metformin induces autophagy and G0/G1 phase cell cycle arrest in myeloma by targeting the AMPK/mTORC1 and mTORC2 pathways.
Metformin induces autophagy and G0/G1 phase cell cycle arrest in myeloma by targeting the AMPK/mTORC1 and mTORC2 pathways.
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二甲双胍通过靶向 AMPK/mTORC1 和 mTORC2 通路诱导骨髓瘤中的自噬和 G0/G1 期细胞周期停滞
DOI：
10.1186/s13046-018-0731-5
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发表时间：
2018-03-20
期刊：
影响因子：
--
通讯作者：
Yan H
中科院分区：
文献类型：
--
作者：
Wang Y;Xu W;Yan Z;Zhao W;Mi J;Li J;Yan H
背景二甲双胍是治疗糖尿病的常用药物。越来越多的证据表明它对多种癌症具有抗肿瘤作用，包括多发性骨髓瘤 (MM)；方法采用人MM细胞系(RPMI8226和U266)体外和体内NOD-SCID小鼠异种移植MM模型评价二甲双胍的抗骨髓瘤作用。使用 CCK8 评估细胞活力，并通过 EdU 掺入测定测量细胞增殖。通过流式细胞术检查细胞周期分布和凋亡。使用透射电子显微镜观察自噬体。通过蛋白质印迹分析评估 AMPK 的激活和 mTORC1/C2 通路的抑制。使用Lipofectamine 2000试剂转染针对AMPK-α1和α2亚基的小干扰RNA，产生AMPK敲低的RPMI8226细胞和U266细胞系。结果二甲双胍有效抑制MM细胞系的增殖，这种作用与诱导自噬和G0/G1细胞周期阻滞有关，但与细胞凋亡无关。二甲双胍激活 AMPK 并抑制骨髓瘤细胞中的 mTORC1 和 mTORC2 信号通路以及下游分子信号通路，例如 p-4EBP1 和 p-AKT。 AMPK 激活导致结节性硬化症复合物 2 (TSC2) 直接磷酸化和激活，从而抑制哺乳动物雷帕霉素靶点 (mTOR)。此外，二甲双胍以 AMPK 依赖性方式抑制骨髓瘤细胞生长。异种移植小鼠模型进一步证实二甲双胍通过上调AMPK和下调mTOR抑制肿瘤生长。结论二甲双胍通过诱导自噬和细胞周期阻滞抑制骨髓瘤细胞的增殖。我们的结果表明，分子机制涉及通过 AMPK 激活对 mTORC1 和 mTORC2 途径的双重抑制。我们的研究为使用二甲双胍作为一种已获批准的安全药物来开发治疗 MM 的新策略提供了理论基础。
BackgroundMetformin is a commonly used drug for the treatment of diabetes. Accumulating evidence suggests that it exerts anti-tumor effects in many cancers, including multiple myeloma (MM); however, the underlying molecular mechanisms have not been clearly elucidated.MethodsThe anti-myeloma effects of metformin were evaluated using human MM cell lines (RPMI8226 and U266) in vitro and in vivo NOD-SCID murine xenograft MM model. Cell viability was assessed with CCK8 and cell proliferation was measured by EdU incorporation assay. Cell cycle distribution and apoptosis were examined by flow cytometry. Transmission electron microscopy was used to visualized autophagosomes. Activation of AMPK and inhibition of mTORC1/C2 pathways was assessed by Western blot analysis. RPMI8226 cells and U266 cell lines with AMPK knockdown were generated by transfection with small interfering RNA targeting the AMPK-α1 and α2 subunits using Lipofectamine 2000 reagent.ResultsMetformin effectively inhibited the proliferation of MM cell lines, an effect that was associated with the induction of autophagy and G0/G1 cell cycle arrest, but not apoptosis. Metformin activated AMPK and repressed both mTORC1 and mTORC2 signaling pathways in myeloma cells as well as downstream molecular signaling pathways, such as p-4EBP1 and p-AKT. AMPK activation resulted in direct phosphorylation and activation of tuberous sclerosis complex 2 (TSC2), leading to inhibition of the mammalian target of rapamycin (mTOR). In addition, metformin inhibited myeloma cell growth in an AMPK-dependent manner. The xenograft mouse model further confirmed that metformin inhibited tumor growth by upregulation of AMPK and downregulation of mTOR.ConclusionsMetformin inhibits the proliferation of myeloma cells by inducing autophagy and cell-cycle arrest. Our results suggest that the molecular mechanism involves dual repression of mTORC1 and mTORC2 pathways via AMPK activation. Our study provides a theoretical basis for the development of novel strategies for the treatment of MM using metformin as an already approved and safe drug.
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影响因子：
29
作者：
Howell JJ;Hellberg K;Turner M;Talbott G;Kolar MJ;Ross DS;Hoxhaj G;Saghatelian A;Shaw RJ;Manning BD
通讯作者：
Manning BD
影响因子：
3.9
作者：
Anisimov, VN;Berstein, LM;Franceschi, C
通讯作者：
Franceschi, C
影响因子：
5.7
作者：
Breuleux M;Klopfenstein M;Stephan C;Doughty CA;Barys L;Maira SM;Kwiatkowski D;Lane HA
通讯作者：
Lane HA
影响因子：
4.8
作者：
Granato, Marisa;Gilardini Montani, Maria Saveria;Cirone, Mara
通讯作者：
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胰腺癌细胞中Berberine和二甲双胍抑制MTORC1，ERK，DNA合成和增殖的剂量依赖性AMPK依赖性和独立机制。
影响因子：
3.7
作者：
Ming M;Sinnett-Smith J;Wang J;Soares HP;Young SH;Eibl G;Rozengurt E
通讯作者：
Rozengurt E