抑制 IGF1R 增强 2-脱氧葡萄糖治疗非小细胞肺癌。
Inhibition of IGF1R enhances 2-deoxyglucose in the treatment of non-small cell lung cancer.
机构信息
Department of Hematology and Medical Oncology, Emory University School of Medicine, Atlanta, GA, 30322, USA; Department of Gastrointestinal Surgery, The First Affiliated Hospital of Sun Yat-sen University, Guangzhou, PR China.
Department of Biostatistics and Bioinformatics, Emory University Rollins School of Public Health, Atlanta, GA, 30322, USA.
出版信息
Lung Cancer. 2018 Sep;123:36-43. doi: 10.1016/j.lungcan.2018.06.026. Epub 2018 Jun 23.
OBJECTIVE
We previously postulated that 2-deoxyglucose (2-DG) activates multiple pro-survival pathways through IGF1R to negate its inhibitory effect on glycolysis. Here, we evaluated whether IGF1R inhibitor synergizes with 2-DG to impede the growth of non-small cell lung cancer (NSCLC).
MATERIALS AND METHODS
The activation of IGF1R signaling was assessed by the phosphorylation of IGF1R and its downstream target AKT using immunoblot. Drug dose response and combination index analyses were carried out according to the method of Chou and Talalay. Flow cytometry was used to evaluate cell cycle progression. Apoptosis was monitored by caspase-3/PARP cleavages or Annexin V staining. A subcutaneous xenograft model was used to assess this combination in vivo.
RESULTS
2-DG induces the phosphorylation of IGF1R in its kinase domain, which can be abolished by the IGF1R inhibitor BMS-754807. Furthermore, the combination of 2-DG and BMS-754807 synergistically inhibited the survival of several non-small cell lung cancer (NSCLC) cell lines both in vitro and in vivo. The mechanistic basis of this synergy was cell line-dependent, and LKB1-inactivated EKVX cells underwent apoptosis following treatment with a subtoxic dose of 2-DG and BMS-754807. For these cells, the restoration of LKB1 kinase activity suppressed apoptosis induced by this combination but enhanced G1 arrest. In H460 cells, the addition of 2-DG did not enhance the low level of apoptosis induced by BMS-754807. However, treatment with 0.75 μM of BMS-754807 resulted in the accumulation of H460 cells with 8n-DNA content without affecting cell density increases. Hence, H460 cells may escape BMS-754807-induced G2/M cell cycle arrest through polyploidy. The inclusion of 2-DG blocked formation of the 8n-DNA cell population and restored G2/M phase cell cycle arrest.
CONCLUSION
The combination of 2-DG and IGF1R inhibitor BMS-754807 may be used to suppress the proliferation of NSCLC tumors through different mechanisms.
目的
我们之前假设 2-脱氧葡萄糖（2-DG）通过 IGF1R 激活多种存活相关途径，从而抵消其对糖酵解的抑制作用。在此，我们评估 IGF1R 抑制剂与 2-DG 联合应用是否能抑制非小细胞肺癌（NSCLC）的生长。
材料与方法
通过 IGF1R 及其下游靶标 AKT 的磷酸化来评估 IGF1R 信号的激活情况，采用免疫印迹法进行检测。根据 Chou 和 Talalay 的方法进行药物剂量反应和合并指数分析。采用流式细胞术评估细胞周期进程。通过 caspase-3/PARP 裂解或 Annexin V 染色监测细胞凋亡。采用皮下移植瘤模型在体内评估该联合用药的效果。
结果
2-DG 诱导 IGF1R 在其激酶结构域磷酸化，这一过程可被 IGF1R 抑制剂 BMS-754807 所阻断。此外，2-DG 与 BMS-754807 联合应用在体外和体内均能协同抑制多种非小细胞肺癌（NSCLC）细胞系的存活。这种协同作用的机制基础具有细胞系依赖性，在 LKB1 失活的 EKVX 细胞中，用亚毒性剂量的 2-DG 和 BMS-754807 处理后会发生细胞凋亡。对于这些细胞，LKB1 激酶活性的恢复会抑制该联合用药诱导的细胞凋亡，但会增强 G1 期阻滞。在 H460 细胞中，2-DG 的添加并不能增强 BMS-754807 诱导的低水平细胞凋亡。然而，用 0.75 μM 的 BMS-754807 处理会导致 H460 细胞积累 8n-DNA 含量而不影响细胞密度增加。因此，H460 细胞可能通过多倍体逃避 BMS-754807 诱导的 G2/M 细胞周期阻滞。2-DG 的加入阻止了 8n-DNA 细胞群的形成，并恢复了 G2/M 期细胞周期阻滞。
结论
2-DG 与 IGF1R 抑制剂 BMS-754807 的联合应用可能通过不同的机制抑制 NSCLC 肿瘤的增殖。
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