Nobiletin Inhibits Non-Small-Cell Lung Cancer by Inactivating WNT/β-Catenin Signaling through Downregulating miR-15-5p

Figure 5

Nobiletin upregulates the negative regulator expression of WNT/β-catenin signaling and miR-15-5p expression. (a) Heatmap showing that nobiletin affected the expression levels of many miRNAs in NSCLC cells. (b) Nobiletin inhibited miR-15-5p expression in both A549 and H460 cells. (c) Predicted binding sites of miR-15-5p in the 3′-UTRs of NKD1, AXIN2, and WIF1. (d) The inhibitory effect of nobiletin on miR-15-5p expression was measured by qRT-PCR in NSCLC cells. (e) Luciferase activity of the reporter driven by the 3′-UTRs of NKD1, AXIN2, and WIF1 in A549 cells cotransfected with negative control oligonucleotides (NC) or miR-15-5p. (f) The transfection efficacy of miR-15-5p was measured by qRT-PCR. (g) The effects of miR-15-5p on the mRNA expression levels of NKD1, AXIN2, and WIF1 were measured by qRT-PCR. (h) The effects of miR-15-5p on the protein expression levels of NKD1, AXIN2, and WIF1 were measured by western blot.