Effect of targeted regulation of the Wnt2 gene by microRNA(microRNA-21) on the proliferation and migration of HepG2 hepatoma cells
10.3969/j.issn.1001-5256.2020.04.019
- VernacularTitle:microRNA-21靶向调控Wnt2基因对肝癌细胞HepG2增殖与迁移的影响
- Author:
Zexin WANG
1
;
Lijun GUAN
;
Jianming LI
Author Information
1. Department of Interventional Radiology, The Affiliated Hospital of Inner Mongolia Medical University, Hohhot 010050, China
- Publication Type:Research Article
- Keywords:
microRNAs;
Wnt2 protein;
carcinoma, hepatocellular;
cell proliferation;
cell movement
- From:
Journal of Clinical Hepatology
2020;36(4):803-807
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo investigate the effect of targeted regulation of the Wnt2 gene by microRNA(miR-21) on the proliferation and migration of HepG2 hepatoma cells. MethodsQuantitative real-time PCR was used to measure the mRNA expression of miR-21 in HepG2 hepatoma cells and normal liver cell line LO2. HepG2 cells were transfected with miR-21 inhibitor, and then the expression of miR-21 and cell proliferation, migration, and apoptosis were analyzed for the inhibitor group and the control group. The protein expression of Wnt2 was measured for the two groups, and dual-luciferase reporter assay was used to verify the association between miR-21 and the Wnt2 gene. The t-test was used for comparison of continuous data between groups. ResultsThe relative expression of miR-21 in HepG2 cells was significantly higher than that in LO2 cells (1.978±0.035 vs 1.586±0.022, t=16.424, P＜0.05). After the transfection of miR-21 inhibitor, the inhibitor group had significantly lower expression of miR-21 than the control group (0.857±0.017 vs 1.684±0.039, t=33669, P＜0.05). Compared with the control group after the transfection of miR-21 inhibitor, the inhibitor group had a significant reduction in the proliferation ability of HepG2 cells (P＜0.05), a significantly lower number of cells passing through the Transwell chamber (83.72±15.06 vs 147.85±20.64, t=4.347, P＜0.05), and a significantly higher cell apoptosis rate (25.67%±3.95% vs 10.27%±2.14%, t=5937, P＜0.05). The inhibitor group had significantly lower relative expression of Wnt2 in HepG2 cells than the control group (0.862±0.127 vs 1.306±0.218, t=3.048, P＜0.05). TargetScan software showed that miR-21 inhibitor significantly inhibited the luciferase activity of the cells transfected with wild-type Wnt2-3′UTR plasmid (0.972±0.102 vs 0.612±0.092, t=4.219, P＜005), while there was no effect on the luciferase activity of the cells transfected with mutant Wnt2-3′UTR plasmid (0.982±0.093 vs 0911±0.128, t=0.972, P＞0.05). ConclusionInhibition of miR-21 expression can effectively inhibit the proliferation and migration of HepG2 cells, promote the apoptosis of HepG2 cells, and inhibit the over-activation of the Wnt signaling pathway, and therefore, it may become one of the potential target genes for liver cancer treatment.
