Negative Pressure Promotes G3BP1-Mediated Migration of Corneal Epithelial Cells Through Activation of AKT/ERK/Paxillin Pathway

The corneal epithelium serves as the outermost transparent barrier of the eye and depends on rapid and coordinated cellular responses for wound repair. Although negative pressure (NP) has been shown to accelerate wound healing in other tissues, its cellular and molecular effects on corneal epithelium remain undefined. This study investigates how NP regulates corneal epithelial cell physiology and identifies the molecular mechanisms underlying NP-induced migration. Human corneal epithelial cells were exposed to normal or NP conditions, and cell motility was quantified using scratch-wound and transwell migration assays. Nuclear and cytoplasmic fractions were isolated for proteomic profiling to identify NP-responsive proteins. G3BP1 was selected as a candidate regulator and subsequently examined using molecular, biochemical, and functional assays to determine its role in NP-mediated signaling. Proteomic analysis revealed a significant NP-induced upregulation of G3BP1. Mechanistically, G3BP1 suppressed epithelial junctional proteins, including E-cadherin, p120-catenin, and ZO-1, while activating key pro-migratory signaling pathways involving AKT, ERK1/2, FAK, and Paxillin. These coordinated changes enhanced cytoskeletal dynamics and promoted corneal epithelial cell migration under NP stimulation. G3BP1 functions as a critical mechanotransduction mediator of NP, orchestrating adhesion remodeling and activating pro-migratory signaling cascades to facilitate corneal epithelial cell motility. These findings reveal a previously unrecognized cellular mechanism through which NP promotes epithelial repair and highlight G3BP1 as a potential therapeutic target for persistent corneal epithelial defects.