Effect of miR-30c targeting Wnt/β-catenin signal on the proliferation and apoptosis of human retinal endothelial cells induced by high glucose / 中华内分泌外科杂志

ABSTRACT

Objective:To investigate the protective effect and mechanism of miR-30c targeting Wnt/β-catenin signal on the proliferation and apoptosis of human retinal endothelial cells (HRECs) induced by high glucose.Methods:Human retinal endothelial cells (HRECs) were cultured and given normal concentration (control group) and high concentration glucose (high glucose group) respectively. According to the experimental design, miR-30c mimic, negative control (miR-NC), Wnt1 overexpression vector (pcDNA3.1-Wnt1) and empty vector (pcDNA3.1) were transfected respectively. RT-PCR was used to detect the expression level of miR-30c in each group. Western blot was used to detect the expression levels of Wnt1, β-catenin and GSK-3β protein in each group. The dual luciferase experiment verified the targeting relationship of miR-30c to Wnt1. Thiazole blue method was used to detect the proliferation activity of each group. Flow cytometry was employed to detect the level of apoptosis of each group of cells.Results:Compared with the control group, the expression of miR-30c in the high glucose group was significantly reduced [ (0.94±0.11) vs (0.32±0.06), P<0.001]; compared with the control group, the cell proliferation activity of the high glucose group was significantly reduced, and the apoptosis rate was significantly increased [ (0.75±0.08) vs (0.13±0.04), (3.53±0.29) % vs (14.89±0.94) %, P<0.001]; compared with the high glucose+miR-NC group, the cell proliferation activity of the high glucose+miR-30c group was significantly increased, and the apoptosis rate was reduced [ (0.14±0.04) vs (0.64±0.06), (14.14±0.86) % vs (6.28±0.45) %, P<0.001]; compared with the miR-NC group, the luciferase activity of the miR-30c group co-transfected with WT-Wnt1 was significantly reduced [ (0.97±0.09) vs (0.26±0.03), P<0.001]; compared with the control group, the protein expression of Wnt1, β-catenin and GSK-3β in the high glucose group were significantly increased [ (0.43±0.05) vs (1.02±0.09), (0.25±0.04) vs (0.82±0.10), (0.39±0.04) vs (0.76±0.11), P<0.001]; compared with the high glucose+miR-NC group, the protein expression of Wnt1, β-catenin and GSK-3β in the high glucose+miR-30c group were significantly reduced [ (1.04±0.10) vs (0.68±0.06), (0.79±0.09) vs (0.34±0.05), (0.74±0.12) vs (0.48±0.06), P<0.001]; compared with the high glucose+miR-30c group, the cell proliferation activity was significantly reduced in the high glucose+miR-30c+pcDNA3.1-Wnt1 group, and the apoptosis rate was significantly increased [ (0.66±0.07) vs (0.31±0.05), (4.26±0.57) % vs (9.75±0.85) %, P<0.001]. Conclusion:miR-30c may negatively regulate the Wnt/β-catenin signaling pathway, promote cell proliferation, and inhibit cell apoptosis induced by high glucose.