Anemoside B4 inhibits non-small cell lung cancer metastasis by modulating epithelial-mesenchymal transition mediated by the MAPK and Keap1/Nrf2 signaling pathways

ObjectiveTo investigate the effect and mechanism of Pulsatilla saponin B4 (AB4) on the metastasis of non-small cell lung cancer by regulating epithelial-mesenchymal transition (EMT).MethodsIn vitro, MTT assay was used to detect the effect of AB4 on the viability of A549 cells. Wound healing and Transwell assays were used to detect the effect of AB4 on the migration and invasion of TGF-β1-induced A549 cells. Western blot and immunofluorescence were used to detect the expression of EMT, MAPK and oxidative stress signaling pathway-related proteins. In vivo, a mouse lung cancer metastasis model was established by tail vein injection of B16-F10 cells. The effect of Pulsatilla saponin B4 (20, 40 mg/kg) on lung metastasis in mice was observed. Meanwhile, blood analyzer was used to detect blood indicators, HE staining was used to examine pathological changes in lung tissue, and Western blot was used to detect the expression of EMT, MAPK and oxidative stress signaling pathway-related proteins in lung tissue.ResultsIn vitro experiments showed that Pulsatilla saponin B4 (5, 10, 20 μmol/L) had no cytotoxicity. Wound healing and Transwell assays showed that Pulsatilla saponin B4 could inhibit the migration and invasion of A549 cells, decrease the expression of N-cadherin, vimentin and transcription factors Slug and Snail in the EMT signaling pathway, and increase the expression of E-cadherin protein; decrease the expression of Keap1 in the oxidative stress signaling pathway and increase the expression of Nrf2; in addition, Pulsatilla saponin B4 could decrease the protein levels of p-JNK, p-ERK and p-p38 in the MAPK signaling pathway (all P