Effect of siRNA on Wisp-1 gene expression, proliferation, migration and adhesion of mouse hepatocellular carcinoma cells

摘要

Objective:To study the inhibition effect of siRNA on the expression of Wisp-1 gene in Hca-F of mouse hepatocellular carcinoma cells strain and also its effect on the proliferation,migration and adhesion of hepatocellular carcinoma cells.Methods:Three expression vectors of siRNA were constructed.Lipo2000 was employed to transfect Hca-F cells and Western blot was used to detect the inhibition effect of siRNA on the expression of Wisp-1 gene.Afterwards,CCK8 was adopted to detect the effect of Wisp-1 siRNA on the proliferation of Hca-F cells;Annexin V-FTTC/PI double staining flow cytometry was used to detect the effect of Wisp-1 siRNA on the apoptosis of Hca-F cells;Transwell was used to detect the effect of Wisp-1 siRNA on the migration of Hca-F cells.The in vitro cell adhesion kit was used to detect of Wisp-1 siRNA on the change in the components of extracellular matrix to which Hca-F cells adhered.Western blot was used to detect the activation of protein kinase B(AKT)/glycogen synthase kinase-3(3pathway and the expression of downstream target protein p53 and matrix metal!oproteinases-2.Results:The siRNA showed interference effect on the expression of Wisp-1 gene.Compared with the control group,after being transfected to cells,Wisp-1 siRNA could significantly inhibit the proliferation,migration and adhesion of Hca-F cells and also promote the cell apoptosis,which was related to the down-regulated phosphorylation of AKT and glycogen synthase kinase-3β and the expression of p53 and matrix metalIoproteinases-2(P<0.05).Conclusions:The inhibition of Wisp-1 expression can reduce the proliferation,migration and adhesion of mouse hepatocellular carcinoma cells,which is related to the AKT/ glycogen synthase kinase-3 β pathway.Wisp-1 gene may be the potential target to cure the hepatocellular carcinoma.