Figure 4.

miR-150-5p Targeting HMGA2 and Wnt-β-Catenin Signaling in NSCLC

(A) Heatmap showing proteins that were downregulated by miR-150-5p by more than 1.5-fold in sphere-formed NSCLC cells. Sphere-formed H1299 or A549 cells were infected with miR-150-5p-expressing lentivirus or an empty vector. After 48 h, the cells were subjected to proteomics analysis. (B) Venn diagram showing substantial overlap of core proteins from H1299 and A549 cell lines. (C) Predicted binding sites of miR-150-5p in the wild-type 3′ UTRs of HMGA2, GSKIP, and β-catenin (CTNNB1). Mutations in these 3′ UTRs are highlighted in red. (D) HMGA2, GSKIP, and β-catenin expressions were significantly downregulated by miR-150-5p. The expression levels of the indicated proteins were measured using immunohistochemistry (IHC). Tissues from the lung metastasis model were generated with sphere-formed H1299 cells that were infected with miR-150-5p-expressing lentivirus or an empty vector. (E) Luciferase activity of reporter with wild-type or mutant 3′ UTRs of HMGA2, GSKIP, and β-catenin in the A549 cells cotransfected with control oligonucleotides or miR-150-5p mimics. (F) Restored HMGA2 and/or β-catenin blocked miR-150-5p-induced inhibition of sphere formation in sphere-formed H1299 cells. Sphere-formed H1299 cells were infected with the indicated gene expression lentivirus for 24 h, and they were then subjected to sphere formation. (G) Restored HMGA2 and/or β-catenin blocked the inhibition of invasion by miR-150-5p in sphere-formed H1299 cells. Sphere-formed H1299 cells were infected with the indicated gene expression lentivirus for 24 h, and they were then subjected to an invasion assay.