Abstract
Purpose :
The presence of a non-keratinized stratified squamous epithelium protects organs that are in close contact with the external environment and regularly exposed to physical abrasion. Here we investigated the contribution of CD147 to promoting the differentiation and establishment of barrier function in human corneal epithelial cells.
Methods :
CRISPR/Cas9-mediated gene editing was used to produce a human corneal epithelial cell line lacking CD147 expression. Transmission and scanning electron microscopy were used to investigate the ultrastructure of differentiated cell cultures. Protein biosynthesis and localization were determined by immunoblotting and immunofluorescence, respectively. The analysis of genes encoding for tight junction proteins was carried out using a human tight junction PCR array. Epithelial cell permeability was evaluated by transepithelial electrical resistance and rose bengal penetrance.
Results :
Cell cultures lacking CD147 change shape and fail to produce a flattened squamous layer on the apical surface. This process is associated with the decreased biosynthesis of MUC16 and involucrin. Expression analysis of genes encoding tight junction proteins indicates that CD147 promotes the physiological expression of occludin and members of the claudin family. Functionally, disruption of CD147 expression leads to increased epithelial cell permeability as evidenced by the decrease in transepithelial electrical resistance and increase in rose bengal flux.
Conclusions :
CD147 plays a distinctive role in maintaining the normal differentiation of stratified squamous corneal epithelium under homeostatic conditions.
This is a 2021 ARVO Annual Meeting abstract.