Regulation mechanism of miR-204 targeted SIRT1 in human lens epithelial cell apoptosis

Abstract

Objective To investigate the effect of miR-204 expression on apoptosis of human lens epithelial cell line (LECs) SRA01/04 in vitro and the mechanism of targeting SIRT1. Methods Hydrogen peroxide (H2O2) was used to construct the cell apoptosis model of human lens epithelial cell line SRA01/04 in vitro. MiR-204 mimic and inhibitor were transiently transfected into SRA01/04 cells using Lipofectamine 2000 to regulate the expression level of miR-204. Flow cytometry was used to detect the cell apoptosis rate in each group SRA01/04 cells. By microRNA databases and recent study researches, we predicted that SIRT1 maybe a potential target gene of miR-204. Quantitative real-time PCR (qRT-PCR) were used to detect the expression level of miR-204 and SIRT1 gene. Results Quantitative real-time PCR results showed that the expression level of miR-204 in cell apoptosis model of SRA01/04 was significantly higher than the normal SRA01/04 cells. Overexpression of miR-204 in SRA01/04 promoted H2O2-induced cell apoptosis, and down-regulated the expression level of SIRT1 gene. While downexpression of miR-204 in SRA01/04 reduced H2O2-induced cell apoptosis, and up-regulated the expression level of SIRT1 gene. Each result was statistically significant (P <0.01). Conclusions The expression level of miR-204 in cell apoptosis model of SRA01/04 is significantly higher than the normal SRA01/04 cells. MiR-204 can regulate the apoptosis of the human lens epithelial cells by targeting SIRT1 gene. Thus, miR-204 may play an important role in the pathogenesis of caracts with lens epithelial cell apoptosis. Key words: miR-204; SIRT1; Cell apoptosis; Cataract