Abstract
Purpose :
Neural precursor cell-expressed developmentally down-regulated protein 4 (NEDD4) is an important E3 ubiquitin ligase which is implicated in multiple biological processes. However, its role in regulating corneal epithelial renewal, remains elusive. In the present study, we investigated the functional role that NEDD4 may play in corneal epithelial wound healing (CEWH) process.
Methods :
Western blotting was performed to detect Nedd4 expression during mouse CEWH. Specific siRNA targeting NEDD4 was transfected into human corneal epithelial cells (HCECs) using Lipofectamine RNAiMAX reagent. MTS, Flow cytometry, and wound-healing assay evaluated the effects of NEDD4 knockdown on cell proliferation, cell cycle progression, and migration, respectively. The effects of NEDD4 knockdown on in vivo wound healing were evaluated using a murine corneal epithelial debridement model by monitoring corneal fluorescein staining.
Results :
Nedd4 expression level was significantly upregulated during mouse CEWH. Knockdown of NEDD4 significantly inhibited HCEC proliferation by inducing cell cycle G2 arrest and suppressed cell migration. Furthermore, Nedd4 knockdown remarkably retarded in vivo CEWH. In addition, downregulation of NEDD4 decreased p-Akt level in HCECs.
Conclusions :
Our results demonstrate that NEDD4-mediated ubiquitination contributes to promoting CEWH through activation of the Akt signaling pathway.
This is a 2020 ARVO Annual Meeting abstract.