Echinacoside inhibits Skp2-induced epithelial-mesenchymal transition and stemness in glioblastoma
Objective To determine the impact of echinacoside(ECH)on the epithelial-mesenchymal transition(EMT)and glioma stem cell characteristics in glioblastoma,and to elucidate related molecular mechanism.Methods ① In vitro experiments:glioblastoma U87 and U251 cells were cultured and treated with 0,5,10,and 20 μmol·L-1 ECH for 24 hours;the cell viability was evaluated with the CCK-8 assay;clonogenic capacity was determined by the colony formation assay,migration and invasion by transwell assay,and stem cell potential by sphere formation assay.The expression of stem cell markers was assessed with immunofluorescence;EMT and stemness-related protein expressions were analyzed with Western blot.② In vivo experiments:U87 cells were subcutaneously implanted into nude mice(five mice in each group).The experimental group was administered daily intraperitoneal injections of ECH,while the control group received a corresponding volume of the vehicle.At the end of the experiment,tumor volume,mouse weight,tumor weight were measured,and Skp2,EMT,and stemness-related protein expressions in tumors were analyzed by Western blot.Results ① In vitro experiments showed that,compared to the 0 μmol·L-1 ECH group,the 10 and 20 μmol·L-1 ECH groups significantly reduced cell viability(P＜0.05);the number of colonies formed was significantly decreased in the 5,10,and 20 μmol·L-1 ECH groups(P＜0.05);migration and invasion abilities were reduced in the 5 and 10 μmol·L-1 ECH groups(P＜0.05);spheroid formation ability and stem cell marker expression,Skp2 and EMT expression,and stem cell characteristic proteins significantly decreased in the 5 and 10 μmol·L-1 ECH groups(P＜0.05).(2)In vivo experiments showed that,tumor volume and weight in mice treated with ECH were obviously smaller than those in the control group(P＜0.05),and the expression of Skp2 and EMT markers(Vimentin,Snail),and stem cell markers(Sox2,Nestin)significantly decreased;there was no significant difference in the body weight between both groups.Conclusion ECH inhibits the EMT and diminishes stemness in glioblastoma by downregulating Skp2 expression.
NETL
NSTL
万方数据
