目的：探究抗烯醇化酶 1（enolase 1，ENO1）抗体、二甲双胍（metformin，MET）通过靶向肿瘤干细胞对西妥昔单抗（cetuximab，CTX）耐药型非小细胞肺癌A549细胞增殖、迁移、侵袭和干性的影响及其可能的作用机制。方法：10 mmol/L MET联合40 μg/ml抗ENO1抗体处理CTX（35 μg/ml）耐药型非小细胞肺癌细胞A549，4 d后通过增殖、克隆形成、迁移、侵袭、甲基纤维素成球实验等检测联合治疗对A549肺癌细胞的影响。同时通过流式细胞术检测CTX、MET和抗ENO1抗体分别以单药及三药联合处理对ALDH+和CD44+两种肺癌干细胞亚群的影响。结果：CTX、MET和抗ENO1抗体联合处理显著抑制A549细胞的增殖、迁移、侵袭能力及A549细胞的自我更新能力。流式细胞术分析发现MET可显著抑制ALDH+干细胞亚群，而抗ENO1抗体可显著抑制CD44+干细胞亚群，三药联合处理可同时显著抑制ALDH+和CD44+干细胞亚群。结论：MET和抗ENO1抗体分别靶向ALDH+和CD44+肿瘤干细胞亚群，两者联合可有效逆转A549细胞对CTX的抗性，从而更有效地抑制A549细胞的干性、增殖、迁移和复发。

Objective: To explore the effect of anti-ENO1 (enolase 1) antibody and metformin (MET) treatment on the proliferation,migration, invasion and stemness of cetuximab (CTX) -resistant non-small cell lung cancer (NSCLC) cells through targeting cancer stem cells and the possible mechanism. Methods: 10 mmol/L MET combined with 40 μg/ml anti-ENO1 antibody was used to treat CTX (35 μg/ml)-resistant NSCLC A549 cells for 4 d, and the effects of combined treatment on A549 cells were detected with proliferation experiment, colony formation assay, migration and invasion experiments and methylcellulose ball formation experiment. In the meanwhile, FCM was used to detect the effects of CTX, MET and anti-ENO1 antibody single-drug treatment as well as the three-drug combination treatment on ALDH+ and CD44+ lung cancer stem cell subsets. Results: CTX combined with MET and anti-ENO1 antibody treatment significantly inhibited the proliferation, migration, invasion and self-renewal capacity of A549 cells. FCM analysis found that MET could significantly inhibit ALDH+ stem cell subpopulations, while anti-ENO1 antibody could significantly inhibit CD44+ stem cell subpopulations, and the three-drug combination treatment could simultaneously suppress ALDH+ and CD44+ stem cell subpopulations. Conclusion: MET and anti-ENO1 antibody respectively target ALDH+ and CD44+ cancer stem cell subsets, and the combined treatment of MET and anti-ENO1 antibody can effectively reverse the resistance of A549 cells to CTX, and thereby more effectively inhibiting stemness, proliferation, metastasis of A549 cells and tumor recurrence.

国家重点研发计划项目（No.2017YFC1308700）；国家自然科学基金面上项目（No.82073278）；中国医学科学院医学与健康科技创新工程项目（No. 2016-I2M-3-013）