Baicalein Inhibits Osteosarcoma Cell Proliferation And Invasion Through The Mir-183/Ezrin Pathway

Objective: Osteosarcoma(OS)is a highly malignant primary bone tumor,baicalein is an effective antitumor drug.The aim of this paper is to study the role of miR-183 /Ezrin signaling pathway in the antiosteosarcoma of baicalein.Methods: Cultivate Saos-2、MG-63 and h FOB cells.Cells were treated with baicalein at different concentrations(25,50,75 and 100 μM),then were cultured for 24,48 or 72 h.Subsequently,the proliferation assay was performed using a CCK-8.After 100 μM baicalein deals with the Saos-2 and MG-63 cells,the cell apoptosis rate was detected by flow cytometry,and the cell migration and invasion ability were detected by Trasnwell.Bioinformatics and biluciferase reporter gene analysis of mir-183 and Ezrin targeting.The expression of Ezrin m RNA and protein was detected by q RT-PCR and western blot after 100 μM baicalein deals with the Saos-2 and MG-63 cells.With miR-183 inhibitor and pc DNA3.1-Ezrin pretreatment of MG-63 for 6 h,then MG-63 and Saos-2 cells confronted by 100 μM baicalein for 48 h.CCK-8 was used to detect Cell proliferation,cell apoptosis rate Transwell migration and invasion assay.Flow cytometry detection of cell apoptosis rate Transwell assay test the cell migration and invasion ability.Results:Baicalein inhibited the proliferation of MG-63 and Saos-2 cells in a dose-and time-dependent manner(P<0.01),the apoptosis rate of MG-63 and Saos-2 cells was low in control group but significantly elevated in baicalein-treated group,After stimulation of MG-63 and Saos-2 cells with 100 μM baicalein for 48 h,the migration and invasion ability of MG-63 and Saos-2 cells was observably impeded.Bioinformatics confirmed that miR-183 and Ezrin 3-’UTR targeted binding sites.Dual luciferase reporter gene detection results show that the luciferase activity of cells transfected with miR-183 mimic was dramatically decreased in wild type,but not mutant.Furthermore,the q RT-PCR and western blot analysis showed that overexpression of miR-183 noteworthily reduced the levels of Ezrin m RNA and protein compared with control group(P<0.01),which were inversed by miR-183 inhibitor,Baicalein can increase the level of miR-183 and down-regulate the expression of Ezrin。MG-63 and Saos-2 cells were co-cultured with baicalein and miR-183 inhibitors or pc DNAEzrin,the results show that miR-183 inhibitor significantly retarded baicalein-induced high apoptosis rate and low proliferation,migration and invasive ability in MG-63 and Saos-2 cells,compared with baicalein group.Conclusion:1)Ezrin is the target gene of miR-183.2)Baicalein can increase the level of miR-183 and downregulate the expression of Ezrin,thereby inducing the apoptosis of OS cells and inhibiting their proliferation and migration.