Fenbendazole inhibits the proliferation,migration and invasion of lung adenocarcinoma cell by activating autophagy
Objective To explore the effects of fenbendazole on the proliferation,migration and invasion of lung adenocarcinoma cells and the relationship between autophagy and migration invasion.Methods A549 and H358 cells were divided into control group and fenbendazole group(1,2.5,5,10,and 20 μmol/L).Cell proliferation level was detected by CCK-8 method and cell counting method,cell migration and invasion ability was detected by scratch assay and Transwell assay,mRNA and protein expression levels of epithelial-mesenchymal transformation(EMT)related proteins were detected by RT-qPCR and western blot.Autophagy related genes and their expression levels were analyzed by transcriptome sequencing.The cells were treated with autophagy inhibitors chloroquine(5,10 μmol/L)and fenbendazole(5 μmol/L),and then the migration and invasion ability of the cells were detected by scratch and Transwell assay.Results Compared with control group,different concentration fenbendazole treatment groups significantly inhibited the activity,migration and invasion ability of A549 and H358 cells(P<0.05,0.01,0.001).With the increase of fenbendazole concentration,the relative mRNA expressions of N-cadherin,Cytokeratin,and FN1 decreased gradually(P<0.05,0.01,0.001).Occludin and Vimentin mRNA expression levels were higher at low concentration,and decreased with the increase of fenbendazole concentration(P<0.05,0.01,0.001).Compared with control group,the relative expression of N-cadherin,Cytokeratin,FN1,and Occludin decreased significantly with the increase of fenbendazole concentration.The relative expression level of Vimentin protein increased with the decrease of drug concentration(P<0.05,0.01,0.001).The thermal map of gene difference showed that the expression of autophagy related genes(MAP1LC3B,ATG5,ULK3,Bax,ATG16L1)increased after fenbendazole treatment,and the expression of autophagy related proteins Beclin-1 and LC3-II/LC3-I increased with the increase of fenbendazole concentration.The expression of P62 showed a decreased trend(P<0.05,0.01,0.001).After chloroquine was used to block autophagy,the inhibitory effects of fenbendazole on cell proliferation,migration and invasion could be reversed(P<0.05,0.01,0.001).Conclusion Fenbendazole can inhibit the proliferation,migration,and invasion abilities of lung adenocarcinoma cells by activating autophagy.
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