Metformin Sensitizes EGFR-TKI-Resistant Human Lung Cancer Cells In Vitro and In Vivo through Inhibition of IL-6 Signaling and EMT Reversal
Metformin Sensitizes EGFR-TKI-Resistant Human Lung Cancer Cells In Vitro and In Vivo through Inhibition of IL-6 Signaling and EMT Reversal
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二甲双胍通过抑制 IL-6 信号传导和 EMT 逆转，在体外和体内使 EGFR-TKI 耐药的人肺癌细胞变得敏感
DOI：
10.1158/1078-0432.ccr-13-2613
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发表时间：
2014-05-15
影响因子：
11.5
通讯作者：
He, Yong
中科院分区：
文献类型：
--
作者：
Li, Li;Han, Rui;He, Yong
目的：EGF受体酪氨酸激酶抑制剂（EGFR-TKI）已成为EGFR激活突变患者的标准治疗方法。不幸的是，获得性耐药最终限制了EGFR-TKI的临床效果和应用。研究表明，抑制上皮-间充质转化（EMT）和白细胞介素（IL）-6/STAT3通路可消除TKI的这种获得性耐药机制。本研究旨在探讨二甲双胍通过抑制IL-6信号传导和逆转EMT在体外和体内对EGFR-TKI耐药人肺癌细胞的增敏作用。实验设计：通过MTT、BrdUrd掺入试验、侵袭试验、流式细胞术分析、免疫染色、Western印迹分析和异种移植物植入，在体外和体内检测二甲双胍逆转TKI耐药的作用。在这项研究中，二甲双胍，一种广泛使用的抗糖尿病药物，有效地增加了TKI耐药肺癌细胞对厄洛替尼或吉非替尼的敏感性。二甲双胍逆转了TKI耐药细胞中的EMT并降低了IL-6信号转导激活，而向这些细胞中加入IL-6则绕过了二甲双胍的抗TKI耐药作用。此外，过表达或添加IL-6至TKI敏感细胞诱导TKI抗性，这可以通过二甲双胍克服。最后，基于二甲双胍的组合疗法有效地阻断了具有TKI抗性癌细胞的异种移植物中的肿瘤生长，这与体内IL-6分泌和表达降低、EMT逆转以及IL-6信号传导激活降低相关。二甲双胍通常被认为是无毒且非常便宜的，可与TKI联合用于非小细胞肺癌患者，携带EGFR突变以克服TKI耐药性并延长生存期。
Purpose: The EGF receptor tyrosine kinase inhibitors (EGFR-TKI) have become a standard therapy in patients with EGFR-activating mutations. Unfortunately, acquired resistance eventually limits the clinical effects and application of EGFR-TKIs. Studies have shown that suppression of epithelial-mesenchymal transition (EMT) and the interleukin (IL)-6/STAT3 pathway may abrogate this acquired mechanism of drug resistance of TKIs. This study aims to investigate the effect of metformin on sensitizing EGFR-TKI-resistant human lung cancer cells in vitro and in vivo through inhibition of IL-6 signaling and EMT reversal.Experimental Design: The effect of metformin on reversing TKI resistance was examined in vitro and in vivo using MTT, BrdUrd incorporation assay, invasion assay, flow cytometry analysis, immunostaining, Western blot analysis, and xenograft implantation.Results: In this study, metformin, a widely used antidiabetic agent, effectively increased the sensitivity of TKI-resistant lung cancer cells to erlotinib or gefitinib. Metformin reversed EMT and decreased IL-6 signaling activation in TKI-resistant cells, while adding IL-6 to those cells bypassed the anti-TKI-resistance effect of metformin. Furthermore, overexpression or addition of IL-6 to TKI-sensitive cells induced TKI resistance, which could be overcome by metformin. Finally, metformin-based combinatorial therapy effectively blocked tumor growth in xenografts with TKI-resistant cancer cells, which was associated with decreased IL-6 secretion and expression, EMT reversal, and decreased IL-6-signaling activation in vivo.Conclusion: Metformin, generally considered nontoxic and remarkably inexpensive, might be used in combination with TKIs in patients with non-small cell lung cancer, harboring EGFR mutations to overcome TKI resistance and prolong survival.