Effect of GSK-3βtargeting RNAi recombinant adenovirus on proliferation of human umbilical vein endothelial cells / 中华内分泌代谢杂志

ABSTRACT

Objective To observe the effect of Wnt/β-catenin pathway on proliferation of human umbilical vein endothelial cell (HUVEC) using the glycogen synthase kinase 3β (GSK-3β)-targeting RNAi recombinant adenovirus vector. Methods Homologous recombination and cloning techniques were used to construct RNAi recombinant adenoviral expressive vectors specific to GSK-3β. Then, the adenovirus plasmids was transfected into HEK 293A cells to produce adenovirus and amplify the adenoviral stock. Plaque forming assay was used to titer the adenoviral stock. The GSK-3β and β-catenin protein expressions were detected by Western blot and immunohistochemistry. The proliferation of HUVEC was detected with MTr assay. Results The RNAi adenovirus vectors specific to GSK-3β were successfully produced with high titer. The expression of GSK-3β protein in HUVEC could be down-regulated efficiently by the RNAi adenovirus, along with increased β-catenin protein expression. The proliferation of HUVEC was significantly increased (P < 0.05 or P < 0.01) after infected with GSK-3β RNAi recombinant adenovirus for 3, 5, 7 days. Conclusion RNAi adenovirus is an important tool that can inhibit the expression of GSK-3β efficiently, along with increased β-catenin protein expression. Up-regulating of the Wnt/β-catenin pathway might play an important role in the proliferation of HUVEC.