Abstract
Purpose :
To determine in vitro the mechanism underlying the ability of mitomycin C to reduce corneal epithelial cell migration and enhance sensory axonal reinnervation in vivo.
Methods :
Telomerase immortalized human corneal limbal epithelial cells and primary human corneal epithelial cells were cultured to 70% confluency and treated with 0.02% mitomycin C for 3 hours; cells were washed, media removed, and cells allowed to condition serum free media for 48 hr. Conditioned media from equal numbers of control or mitomycin C treated primary and immortalized human corneal limbal epithelial cells were used in cytokine array studies which were repeated twice. In addition, cell migration and adhesion assays were performed.
Results :
Cytokine arrays show increased expression of IL1ra, IL1a, IL1b, IL6, IL8, G-CSF, GM-CSF, and CXCL1 but no change in the expression of MIF and PAI1 (serpine1) in conditioned media from mitomycin C treated human corneal limbal epithelial cells compared to conditioned media from controls. None of the cytokines assessed were decreased in expression. Human corneal limbal epithelial cells decrease both their migration rate and adhesion when treated with conditioned media derived from mitomycin C treated corneal epithelial cells.
Conclusions :
These data show that transient mitomycin C treatment of corneal epithelial cells induces a gene expression program that leads to both reduced adhesion and cell migration.
This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.