奥利司他联合阿美替尼对肺腺癌H1975细胞增殖、迁移和侵袭的影响

The effect of orlistat combined with almonertinib on the proliferation,migration and invasion of lung adenocarcinoma H1975 cells

目的:探究奥利司他(orlistat,Orli)联合阿美替尼(almonertinib,Alm)对肺腺癌H1975细胞增殖、迁移和侵袭的影响及其机制。方法:不同浓度的Orli、Alm单药及联合用药处理肺腺癌H1975细胞后,CCK-8法检测细胞增殖活性,计算Alm的半数抑制浓度(IC50)及联合指数(CI)。Transwell实验检测细胞的迁移及侵袭能力。免疫印迹法检测细胞内FASN蛋白、EGFR-AKT信号通路相关蛋白、上皮-间质转化因子E-cadherin、N-cadherin的表达水平。结果:与对照组相比,不同浓度的Orli、Alm及联合用药处理后均可在一定程度上抑制H1975细胞的增殖活性,且低剂量Orli联合Alm对H1975细胞增殖活性的抑制具有协同作用(CI<1)。此外,低剂量联合用药组的细胞迁移、侵袭数目较对照组及各单药组均显著减少(P<0.05)。低剂量联合用药组细胞中EGFR、p-EGFR和p-AKT,FASN蛋白表达水平较各组均明显降低(P<0.05);上皮-间质转化因子E-cadherin表达水平上调,N-cadherin表达水平下调(P<0.05)。结论:Orli联合Alm可协同抑制H1975细胞增殖、迁移及侵袭,其作用机制可能与下调FASN蛋白表达、抑制EGFR通路信号分子磷酸化有关。

Objective:To investigate the effect of orlistat(Orli)combined with almonertinib(Alm)on the proliferation,migration and invasion of lung adenocarcinoma H1975 cells and its mechanism.Methods:After treating lung adenocarcinoma H1975 cells with different concentrations of Orli and Alm alone and in combination,cell proliferation activity was detected by CCK-8 assay,and the half inhibitory concentration(IC 50)and combination index(CI)of Alm were calculated.Transwell assay was performed to detect the ability of the cells to migrate and invade.The expression levels of cellular FASN proteins,EGFR-AKT signalling pathway-related proteins,epithelial-mesenchymal transition factors E-cadherin and N-cadherin were detected by Western blotting.Results:Compared with the control group,the proliferative activity of H1975 cells could be inhibited to a certain extent after treatment with different concentrations of Orli,Alm and the combination of drugs,and the inhibition of the proliferative activity of H1975 cells by low-dose Orli combined with Alm had a synergistic effect(CI<1).In addition,the number of cell migration and invasion was significantly reduced in the low-dose combination group compared with the control group and each single-drug group(P<0.05).The expression levels of EGFR,p-EGFR and p-AKT,and FASN proteins in the cells of the low-dose combination group were significantly reduced compared with those of all groups(P<0.05),and the expression levels of the epithelial-mesenchymal transition factor E-cadherin were up-regulated,and the expression levels of N-cadherin were down-regulated(P<0.05).Conclusion:Orli combined with Alm synergistically inhibited the proliferation,migration and invasion of H1975 cells,and its mechanism of action may be related to the down-regulation of FASN protein expression and the inhibition of phosphorylation of EGFR pathway signalling molecules.