Figure 2.

miR-424 and miR-503 directly target WEE1 in ovarian cancer cells

(A and B) WEE1 protein expression after AGO2 siRNA transfection in SKOV3 cells. (C) Mature miR-424 and miR-503 expression in SKOV3 and OVCAR8 adherent cells and spheroids. (D) Linear correlation between miR-424 and miR-503 expression. Inverse correlation between WEE1 mRNA expression and miR-424/miR-503 expression in ovarian tumor tissues. Relationships between variables were determined by the Pearson correlation coefficient. WEE1 protein (E and F) and mRNA (G) expression in response to the overexpression of miR-424, miR-503, or both (miR-424/503) in primary ovarian cancer cells and SKOV3 cells. (H–J) WEE1 protein and mRNA expression after transfection with anti-miR-424/503 in SKOV3 and OVCAR8 cells. (K) Targeting of the WEE1 3′ untranslated region (UTR) via miR-424 and miR-503 overexpression in SKOV3 cells. Luciferase activity data for constructs with the wild-type and mutant 3′ UTR constructs are shown. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001 compared with controls by unpaired two-tailed Student’s t test or one-way analysis of variance (ANOVA) with Bonferroni’s multiple comparison test. Error bars, standard error of the mean.