Amlotinib inhibits cell proliferation,migration,invasion and promotes cell apoptosis in non-small cell lung cancer through nuclear factor-KB signaling pathway
Objective To investigate the effects of Anlotinib on the proliferation,migration,inva-sion,and apoptosis of non-small cell lung cancer(NSCLC)cells and explore its possible molecular mecha-nisms.Methods Human NSCLC cell line A549 was cultured in vitro and randomly divided into a drug blank group[0.1％dimethyl sulfoxide(DMSO)]and Anlotinib low-,medium-,and high-dose groups(2.5,5,and 10 µmol/L,respectively).Each group was treated with the corresponding solution and cul-tured for 24 hours.Cell proliferation was detected using the methylthiazolyldiphenyl-tetrazolium bromide(MTT)assay,Cell Counting Kit-8(CCK-8),and 5-Ethynyl-2'-deoxyuridine(EdU)staining assay.Cell migration,invasion,and apoptosis were assessed using scratch assays,Transwell chamber invasion assays,and terminal-deoxynucleotidyl transferase mediated nick end labeling(TUNEL)staining,respectively.Western blotting was performed to measure the expression of apoptosis-related proteins,including B lym-phoma-associated X protein(bax),B lymphoma-2 protein(bcl-2),phosphorylated IκBβ(p-IκB),IκB,and phosphorylated nuclear factor-κB p65 subunit(p-NF-κB p65).A549 cells were treated with NF-κB pathway agonists,followed by the same proliferation,migration,invasion,and apoptosis assays.Results Compared with the drug blank group,the high-dose Anlotinib group exhibited significantly reduced prolifer-ation,migration,and invasion abilities(83.41±2.83 vs.29.87±5.31,0.47±0.02 vs.0.98±0.03,0.54±0.02 vs.1.01±0.03,0.29±0.02 vs.0.97±0.02,21.67±3.06 vs.143.70±3.51,t=15.42,25.74,24.30,43.56,45.50,all P＜0.05)and lower expression levels of p-IκB and p-NF-κB p65,as well as reduced p-IκB/IκB and p-NF-κB-p65/NF-κB-p65 ratios(0.52±0.05 vs.1.02±0.04,0.49±0.04 vs.1.02±0.03,0.08±0.01 vs.0.98±0.06,0.24±0.03 vs.0.92±0.05,t=15.36,18.48,27.25,22.32,all P＜0.05).The apoptosis rate and expression levels of bax and bcl-2 were sig-nificantly higher(2.46±0.09 vs.0.94±0.04,2.35±0.09 vs.0.91±0.06,t=27.09,23.44,all P＜0.05),demonstrating a clear dose-dependent effect.Compared with the high-dose Anlotinib+DMSO group,the high-dose Anlotinib+NF-κB agonist group exhibited significantly reduced proliferation,migra-tion,and invasion abilities,and a significantly increased apoptosis rate(0.96±0.03 vs.0.49±0.03,0.57±0.03 vs.0.26±0.03,91.00±3.61 vs.61.67±4.73,0.96±0.03 vs.1.39±0.08,t=18.89,12.58,8.55,9.20,all P＜0.05).Conclusion Anlotinib inhibits the proliferation,migration,and in-vasion of NSCLC cells and promotes apoptosis,potentially through the inhibition of the NF-κB signaling pathway.
NETL
NSTL
万方数据
