Research Paper Volume 12, Issue 20 pp 20366—20379

Figure 2. miR-140-3p downregulates FOXQ1 expression and suppresses BCa cell proliferation and invasion. (A) Overexpression of miR-140-3p in T24 and UMUC3 cells to determine its effect on FOXQ1 expression by western blot assay. Approximately 50 ug of protein was loaded into each lane 2 to 4 days after transfection. (B) miR-140-3p expression in 30 paired human primary BCa and adjacent normal bladder tissues. (C) Correlation analysis of miR-140-3p and FOXQ1 mRNA level was performed using the Pearson correlation coefficient. (D) Representative immunohistochemical staining of FOXQ1 in BCa and nonmalignant bladder tissue samples. (E) Analysis of microarray sequencing from NCBI GEO Datasets (GSE40355, GPL8227) showed miR-140-3p expression in 16 BCa and 8 nonmalignant bladder tissue samples. (F) OS curves of BCa patients were analyzed according to miR-140-3p expression. Data were download from TCGA. (G) T24 cells were transfected with NC and miR-140-3p mimics. (H) UMUC3 cells were transfected with miR-140-3p inhibitor. Cell growth was measured by MTT assay. (I, J) Transwell invasion assays were performed by transfecting T24 cells with a negative control (NC) or miR-140-3p mimics (I) and UMUC3 cells with NC or miR-140-3p inhibitor (J). The invaded cells were counted in 10 randomly chosen microscopic fields (100X) of each experiment and pooled. (G–J) Each sample was run in triplicate and used in multiple experiments to determine the mean ± SD. *P < 0.05; **P < 0.01 compared to controls.