埃克替尼联合二甲双胍对耐药非小细胞肺癌H1975细胞的抗肿瘤作用  

Anti-tumor effect of icotinib combined with metformin on non-small cell lung cancer H1975cells

作　　者：吴婷婷 杨帆 常珂 任春霞 余自成 WU Ting-ting;YANG Fan;CHANG Ke;REN Chun-xia;YU Zi-cheng(School of Medicine,Tongji University,SHANGHAI 200092,China;Department of Pharmacy,Yangpu Hospital,School of Medicine,Tongji University,SHANGHAI 200090,China)

机构地区：[1]同济大学医学院,上海200092 [2]同济大学附属杨浦医院药学部,上海200090

出　　处：《中国新药与临床杂志》2024年第8期600-606,共7页Chinese Journal of New Drugs and Clinical Remedies

基　　金：上海市科学技术委员会科研计划项目(19ZR1450600);上海市临床药学重点专科建设项目支持课题。

摘　　要：目的研究埃克替尼联合二甲双胍对耐药非小细胞肺癌H1975细胞的抗肿瘤作用及可能机制。方法用CCK-8法检测埃克替尼(0、0.1、1、5、10、20、40μmol·mL^(-1))、二甲双胍(0、2、4、8、16、32、64 mmol·mL^(-1))以及埃克替尼[0、0.125、0.25、0.5、1倍半数抑制浓度(IC50)]联合二甲双胍(0.5倍IC50)对H1975细胞增殖的抑制作用和联合指数(CI);采用划痕实验检测细胞的迁移能力;采用侵袭实验检测细胞的侵袭能力;采用Annexin V-FITC/PI流式细胞术检测细胞凋亡率;采用Western blot法检测相关蛋白表达水平。结果埃克替尼和二甲双胍抑制H1975细胞48 h的IC50分别为(49.90±4.84)μmol·mL^(-1)和(13.20±1.27)mmol·mL^(-1)。埃克替尼与二甲双胍对H1975细胞具有协同抑制作用(CI<1)。与二甲双胍组和埃克替尼组比较,联合用药组的细胞迁移率显著下降、细胞侵袭能力显著降低、细胞凋亡率显著增加(P<0.05);Akt、m TOR的磷酸化蛋白水平和Bcl-2的表达水平显著下降,AMPK的磷酸化蛋白水平和Bax的表达水平显著升高(P<0.05)。结论埃克替尼联用二甲双胍对非小细胞肺癌H1975细胞具有显著的协同抑制增殖作用和促凋亡作用,其机制可能与激活AMPK、抑制Akt/m TOR信号通路有关。AIM To study the anti-tumor effect of icotinib combined with metformin on non-small cell lung cancer(NSCLC)H1975 cells and its possible mechanism.METHODS CCK-8 assay was used to detect the proliferation inhibitory effect and combination index(CI)of icotinib monotherapy(0,0.1,1,5,10,20,40μmol·mL^(-1)),metformin monotherapy(0,2,4,8,16,32,64 mmol·mL^(-1)),and icotinib(0,0.125,0.25,0.5,1 times of IC50)combined with metformin(0.5 times of IC_(50))on the H1975 cells.The migratory ability of the cells was detected by scratch test,and the invasive ability of the cells was detected by invasion test.Annexin V-FITC/PI flow cytometry was used to detect the rate of apoptosis.Western blot was used to detect the expression level of related proteins.RESULTS The IC50 of icotinib and metformin for the inhibition on H1975 cells after 48 h were(49.90±4.84)μmol·mL^(-1)and(13.20±1.27)mmol·mL^(-1),respectively.Icotinib and metformin produced a synergistic effect in H1975 cells(CI<1).Compared with the metformin group and the icotinib group,the migration rate and invasion ability of combination group were significantly decreased,and the apoptosis rate was significantly increased(P<0.05);the expression of p-Akt,p-mTOR and Bcl-2 were significantly decreased,while the expression of p-AMPK and Bax were significantly increased(P<0.05).CONCLUSION Icotinib and metformin have significant synergic antitumor effects and proapoptotic effects on non-small cell lung cancer H1975 cells,and the underlying mechanism may be related to enhancing activation of AMPK and inhibition of Akt/mTOR signaling pathway.

关 键 词：埃克替尼 二甲双胍 癌 非小细胞肺 药物协同作用 

分 类 号：R965[医药卫生—药理学]