ARHGAP10 Is a Direct Downstream Target of miR-3174 and Acts in a Tumor-Suppressor Role in GC
(A–D) The mRNA expression of CLMP (A), ARHGAP10 (B), ZNF471 (C), and LODC1 (D) and in 100 paired GC and adjacent normal tissues were detected using RT-PCR. (E–H) Correlations between miR-3174 and these four genes (E, CLMP; F, ARHGAP10; G, ZNF471; H, LODC1) were measured with Pearson correlation analysis. (I) The mRNA expression of these four genes in MKN45 cells with or without miR-3174 reconstitution and in BGC823 cells with or without miR-3174 inhibition was also determined with RT-PCR. (J) The protein levels of ZNF471, LODC1, CLMP, and ARHGAP10 were verified by western blotting. (K and L) Dual-luciferase reporter assays with wild-type or mutant-type 3′ UTRs of these four genes were performed in BGC823 (K) and MKN45 (L) cells with or without reconstitution of miR-3174, and fluorescence was quantified and converted into histograms. (M) Cell viability was detected with a CCK-8 kit in MKN45 cells with or without suppression of ZNF471, LDOC1, CLMP, and ARHGAP10. β-actin was used as an internal control. Graph represents mean ± SEM; *p < 0.05, **p < 0.01, and ***p < 0.001.