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

OBJECTIVES

To investigate the effect and mechanism of Anemoside B4 (AB4) on non-small cell lung cancer (NSCLC) metastasis.

METHODS

In vitro, the MTT assay was used to evaluate the effect of AB4 on the viability of human NSCLC cell lines A549 and H1975. Cell scratch and Transwell assays were performed to assess the effect of AB4 on the migration and invasion of A549 and H1975 cells induced by transforming growth factor-β1 (TGF-β1). Western blotting and immunofluorescence were used to detect the expression of proteins related to epithelial-mesenchymal transition (EMT), the mitogen-activated protein kinase (MAPK) signaling pathway, and the oxidative stress signaling pathway. Invivo, a mouse model of melanoma lung metastasis was established by tail vein injection of B16-F10 cells to evaluate the effect of AB4 (20, 40 mg/kg) on melanoma lung metastasis. Blood routine parameters were measured, pathological changes in lung tissue were observed by hematoxylin and eosin staining, and the expression of EMT-, MAPK-, and oxidative stress signaling pathway-related proteins in lung tissue was analyzed by Western blotting.

RESULTS

In vitro, the MTT assay showed that AB4 (5, 10, 20 μmol/L) had no cytotoxic effect. AB4 inhibited the migration and invasion of A549 and H1975 cells induced by TGF-β1; decreased the expression of N-cadherin, vimentin, Slug, and Snail while increasing E-cadherin expression in the EMT pathway; decreased Keap1 expression and increased Nrf2 expression in the oxidative stress pathway; and reduced the phosphorylation levels of JNK, ERK, and p38 in the MAPK pathway. Invivo, AB4 alleviated weight loss, inhibited melanoma lung metastasis, and the mechanism may be related to the inhibition of EMT, oxidative stress, and the MAPK signaling pathway in the lung tissue of model mice.

CONCLUSIONS

AB4 inhibits tumor metastasis by modulating EMT mediated by the MAPK and Keap1/Nrf2 signaling pathways.