Abstract
Purpose:
Wnt5a is a non-canonical Wnt ligand that activates β-catenin-independent pathways for regulation of variety cellular functions, such as cell migration and polarity, that play crucial roles in wound repair. Investigation of Wnt5a signaling may identify therapeutic targets for wound healing in human corneal endothelial cells (CECs).
Methods:
Expression of Wnt5a, Cdc42, Rac1, LIM domain kinase 2 (LIMK2), slingshot 1 (SSH1), and cofilin were analyzed by immunoblotting. Scratch-induced directional migration assay was employed to measure migratory rates. Activation of Cdc42 and Rac1 were determined by GTP pull-down assay. RhoA activity was measured using RhoA specific G-LISA assay kit.
Results:
Wnt5a induced by IL-1β-stimulation activates Cdc42 and subsequently inhibits of RhoA. Inhibition of RhoA leads to parallel dephosphorylation of LIMK2 and SSH1, resulting in dephosphorylation and activation of cofilin leading to enhanced cell migration.
Conclusions:
These findings suggest that Wnt5a enhances cell migration through regulation of cofilin via LIMK2 and SSH1 regulated by Cdc42 and RhoA in human CECs.
Keywords: 481 cornea: endothelium •
765 wound healing •
714 signal transduction