Hypaconitine inhibits TGF-β1-induced epithelial-mesenchymal transition and suppresses adhesion, migration, and invasion of lung cancer A549 cells

摘要

Epithelial-mesenchymal transition (EMT) has been implicated in tumor invasion and metastasis and provides novel strategies for cancer therapy.Hypaconitine (HpA),a diester-diterpenoid alkaloid isolated from the root of the Aconitum species,exhibits anti-inflammatory,analgesic,and especially,cardiotoxic activities.Here,we reported the anti-metastatic potentials of HpA in transforming growth factor-β1 (TGF-β1)-induced EMT in ltmg cancer A549 cells.The cytotoxic effect of HpA was determined by MTT assay.A549 cells were treated with TGF-β1 with or without HpA co-treatment,and the morphological alterations were observed with a microscopy.The expression of E-cadherin,N-cadherin,and NF-κB was determined by both Westem blotting and immunofluorescence analyses.The adhesion,migration,and invasion were detected with Matrigel,wound-healing,and transwell assays,respectively.The expression of Snail was determined by Western blotting.The expression of NF-κB p65,IκBα,and p-IκBα in nuclear and cytosolic extracts was assessed by Western blotting.The results showed that low concentration of HpA (＜16 μmol·L-1) had no obvious cytotoxicity to A549 cells.Morphologically,TGF-β1 treatment induced spindle-shaped alteration in the cells.The upregulation of N-cadherin,NF-κB,and Snail and the downregulation of E-cadherin were detected after TGF-β1 treatment.The adhesion,migration and invasion abilities were also increased by TGF-β1.Besides,TGF-βl induced expression of Snail in a time-dependent manner.Furthermore,TGF-βl induced nuclear translocation of NF-κB p65.All these alterations were dramatically inhibited by HpA co-treatment.In addition,the NF-κB inhibitor PDTC showed similar inhibitory effect.In conclusion,these results showed that HpA inhibited TGF-β1-induced EMT in A549 cells,which was possibly mediated by the inactivation of the NF-κB signaling pathway,providing an evidence for anti-cancer effect of HpA.