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.
ABSTRACT
Objective To detect the expressions of Wnt2 and dishevelled (Dvl) protein in esophageal squamous carci-noma, and analyze their relationship with the occurrence and development of esophageal squamous cell carcinoma. Meth-ods The expression levels of Wnt2 and Dvl protein were detected by Western blot assay in 60 samples of esophageal carci-noma and adjacent non-carcinomatous esophageal tissues, and their relationship with clinical pathological features were ana-lyzed. Results The relative expression levels of Wnt2 and Dvl protein were higher in esophageal squamous carcinoma tis-sue (0.512 ± 0.406, 1.218±1.082) than those of esophageal tissue adjacent to carcinoma (0.153 ± 0.189, 0.505±0.358). There were significant differences in the expression levels of Wnt2 and Dvl protein between different infiltration depth, different TNM stages, and lymph node metastasis (P<0.01). There was a positive correlation between Wnt2 and Dvl protein in esopha-geal squamous carcinoma (r=0.718, P<0.01). Conclusion The high expression levels of Wnt2 and Dvl protein promote the development and metastasis of esophageal squamous cell carcinomas collaboratively via Wnt2 signal transduction path-ways.
ABSTRACT
Objective: To investiget the expression of Wnt2 in human breast cancer tissues and its effect on the invasion of breast cancer cells. Methods: The serum Wnt2 content of patients with breast cancer and the healthy volunteers was detected by ELISA. The expressions of Wnt2 in breast cancer tissues and their adjacent para-cancerous tissues were observed by immunohistochemistry. The primary cells from the breast cancer cells with high- and low-expression of Wnt2 were cultured. The capacities of invasion and adhesion of the breast cancer cells with high- and low-expression of Wnt2 were determined by Transwell assay and cell adhesion experiment, respectively. Results: The serum Wnt2 content of patients with breast cancer was higher than that of the healthy volunteers (P < 0.05). The positive expression rate of Wnt2 in breast cancer tissues was higher than that in the adjacent paracancerous tissues (P < 0.01). The capacities of invasion and adhesion of breast cancer cells with high-expression of Wnt2 were higher than those of the cells with low-expression of Wnt2 (all P < 0.05). Conclusion: The expression level of Wnt2 is higher in breast cancer. The invasion capacity of the breast cancer cells with high-expression of Wnt2 is relatively stronger.