Mechanism of wangzaozin A, an ent-kaurane diterpene, on cytoskeletal rearrangement and migration inhibition of A549 cells / 中草药

ABSTRACT

Objective: To investigate the mechanism of cytoskeletal recombination and migration inhibition induced by wangzaozin A, ent-kaurane diterpenoid, in A549 cells. Methods: The effects of wangzaozin A on cytotoxicity, cell morphology, cytoskeleton and protein expression as well as cell migration were detected in A549 cells by using MTT, microscope observation, Western blotting, immunofluorescence assay and scratch assay. Results: Wangzaozin A induced significant changes in cell morphology at 24, 48 and 72 h, including increased pseudopods, stretched pseudopods and flattened nucleus in A549 cells. Moreover, microtubules and keratin fibers networks in A549 cells also showed obvious rearrangement, which indicated the cytoskeleton had gone through a continuous recombination process. Further, wangzaozin A significantly increased the phosphorylation of extracellular regulated protein kinase (ERK), microtubule-associated protein 4 (MAP4), keratin 8 (K8) (P < 0.05, 0.01), while wangzaozin A-induced phosphorylation of MAP4 and K8 were suppressed in A549 cells treated with ERK inhibitors U0126 (P < 0.05, 0.01); Wangzaozin A inhibited the migration of A549 cells with a correlation between concentration and time. Conclusion: Wangzaozin A can upregulate the phosphorylation of MAP4 and K8 by activating ERK signaling pathway, which can significantly increase the dynamics of MTs and KFs, disturb the dynamic balance of the cytoskeleton, and inhibit the migration of A549 cells.