Abstract
Purpose :
Downregulation of microRNA-145 (miRNA-145) has been correlated with increased severity of pterygium. We hypothesized that forced expression of miRNA-145 could attenuate pterygium growth.
Methods :
Ectopic expression of miRNA-145 in primary pterygium cells was achieved by transfection of miRNA-145 mimics. Cell properties of miRNA-145-transfected pterygium cells were analyzed by MTT assay, migration assay and flow cytometry. The expression of miRNA-145-related targets, mouse double minute 2 (MDM2) and p53, was determined by flow cytometry and immunoblotting.
Results :
Pterygium cells transfected with miRNA-145 mimics showed lower cell viability (5.71 folds, p<0.001) and retarded migration (71.78% vs 82.10%, p<0.05), compared to those transfected with scramble control. Moreover, the proportion of sub-G1 phase in miRNA-145-transfected cells (40.9%) was higher than that in scramble control (7.32%, p < 0.05). Flow cytometry and immunoblotting analyses confirmed that MDM2 expression was reduced in miRNA-145-transfected pterygium cells compared to scramble control. In contrast, the expression of p53, which is negatively regulated by MDM2, was increased after miRNA-145 transfection.
Conclusions :
This study demonstrated that ectopic expression miRNA-145 reduces pterygium cell viability and migration as well as induce cell apoptosis. MDM2 was downregulated, whereas p53 was upregulated by miRNA-145.. Our results suggest that overexpression of miRNA-145 in pterygium cells could be a potential therapeutic strategy for pterygium treatment through MDM2/p53-induced apoptotic pathway.
This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.