Fig. 5

MEG3 suppresses the ubiquitination of p53 by downregulating MDM2 to regulate miR-149-3p/FOXP3 axis in EC cells. A MEG3 expression in cancer and paracancerous tissues detected by RT-qPCR normalized to GAPDH (n = 56). B MDM2 expression in cancer and paracancerous tissues detected by RT-qPCR normalized to GAPDH (n = 56). * p < 0.05 vs. paracancerous tissues. C The correlation between MEG3 and MDM2 expression in EC tissues. D MEG3 and MDM2 expression in AKR cells detected by RT-qPCR normalized to GAPDH. * p < 0.05 vs. control cells. E mRNA expression of MEG3 and MDM2 in AKR cells after overexpressing MEG3 detected by RT-qPCR normalized to GAPDH. * p < 0.05 vs. AKR cells treated with oe-NC. F Silencing efficiency of sh-MDM2 in AKR cells determined by RT-qPCR normalized to GAPDH. G Silencing efficiency of sh-p53 in AKR cells determined by RT-qPCR normalized to GAPDH. * p < 0.05 vs. AKR cells transfected with sh-NC. H The binding relationship between MDM2 and p53 determined by Co-IP. * p < 0.05 vs. IgG. I Effect of MDM2 inhibition on the stability of p53 protein in AKR cells after cycloheximide (CHX, 20 μM) treatment. J p53 expression in AKR cells after silencing MDM2 determined by western blot analysis normalized to GAPDH. K Western blot analysis result of the ubiquitination of p53 in AKR cells with MDM2 inhibition normalized to GAPDH. * p < 0.05 vs. AKR cells transfected with sh-NC. L, Effect of MEG3 overexpression on the stability of p53 protein in AKR cells after cycloheximide (CHX, 20 μM) treatment. M p53 expression in overexpressed MEG3 AKR cells determined by western blot analysis normalized to GAPDH. N Western blot analysis result of the ubiquitination of p53 in AKR cells with MDM2 overexpression normalized to GAPDH. * p < 0.05 vs. AKR cells transfected with oe-NC. O MEG3, p53, miR-149-3p and FOXP3 expression in different AKR cells measured by RT-qPCR normalized to GAPDH and U6. P p53 and FOXP3 expression in different AKR cells measured by western blot analysis normalized to GAPDH. * p < 0.05 vs. AKR cells treated with oe-NC + sh-NC. # p < 0.05 vs. AKR cells treated with oe-MEG3 + sh-NC. Measurement data were expressed as mean ± standard deviation. Data between cancer and paracancerous tissues were compared by paired t test, and data between other two groups were compared by independent sample t test. Comparisons among multiple groups were performed using one way analysis of variance (ANOVA), followed by Tukey's post hoc test, and data at different time points among multiple groups were compared by repeated measures ANOVA, followed by Bonferroni post hoc test. The cell experiment was repeated 3 times