Fig. 5

RUVBL1 is a direct target of miR-370-3p and an oncogene in OS involved in Wnt/β-catenin signaling. a Target genes of miR-370-3p were predicted by TargetScan and compared with differentially expressed genes in the GEO dataset GSE28424. Overlapped genes matching the condition where |fold change| > 1 and p value < 0.0005 were chosen. b The heat map showed the 4 differentially downregulated and 5 upregulated target genes of miR-370-3p. c The upregulated 5 genes were subjected to qRT-PCR in MG63 cells and U2OS cells transfected with either Si2-circMYO10 and miR-370-3p mimics. Data represents the mean ± SD (n = 9). d-e The abundance of RUVBL1 in 10 paired chondroma (n = 10), osteosarcoma tissues (n = 10), hFOB1.19 and osteosarcoma cell lines was detected by qRT-PCR. d Data represents the mean ± SD (n = 90). e Data represents the mean ± SD (n = 9). f The putative binding site of the RUVBL1 3′ UTR for miR-370-3p. g The luciferase reporter plasmids containing wild type or mutated RUVBL1 3′ UTRs were con-transfected with miR-370-3p mimics into 293 T cells. Data represents the mean ± SD (n = 9). i-j The expression of RUVBL1 in MG63 cells transfected with control or miR-370-3p mimics or miR-370-3p inhibitors was detected by h qRT-PCR, i western blot and j immunofluorescence analysis. h Data represents the mean ± SD (n = 9). k RUVBL1 knockdown inhibited the colony formation ability of both MG63 and U2OS cells. Data represents the mean ± SD (n = 3). l Compromised migration and invasion ability were detected in MG63 and U2OS cells with RUVBL1 inhibited. Scale bars = 200 μm. Data represents the mean ± SD (n = 3). m Western blot analysis indicating the inhibited Wnt/β-catenin signaling as evidenced by the downregulation of C-myc, CyclinD1, β-catenin, nuclear β-catenin, N-cadherin and Vimentin with E-cadherin upregulated. Three independent assays were performed in the above assays. c-e, g-h * P < 0.05, ** P < 0.01, *** P < 0.001 (Student’s t-test)