Figure 1.
miR-182-5p expression and association with clinical parameters in bladder cancer tissues.
A. miR-182-5p expression in clinical samples and bladder cancer cell lines, B. Association of miR-182-5p with clinic-pathological parameters, C. Kaplan Meier plots of overall survival.
Figure 2.
Effect of miR-182-5p over-expression on bladder cancer cell function (T24, UM-UC-3).
Two bladder cancer cell lines (T24 and UM-UC-3) were transiently transfected with either miR-182-5p precursor or control (miR-NC). A. Relative miR-182-5p expression, B. Cell viability assay, C. Invasion assay, D. Wound healing assay (24 hours), E. Flow cytometric analysis of apoptosis in miR-NC or miR-182-5p transfected BC cells.
Figure 3.
miR-182-5p binds to the 3′ UTR of RECK and Smad4 mRNAs and down-regulates expression.
A. RECK and Smad4 3′UTR position and complementary miR-182-5p sequences. B. 3′UTR Luciferase assay (miR-NC and miR-182-5p precursor), C. RECK, Smad4 and beta-tubulin protein expression in miR-NC inhibitor or miR-182-5p inhibitor transfected bladder cancer cells (T24, UM-UC-3).
Figure 4.
Effect of RECK and Smad4 over-expression on bladder cancer cell (T24) function.
A. At 24 hours after transfection of either pCMV6-empty, pCMV6-RECK or pCMV6-Smad4 into bladder cancer cells (T24), RECK and Smad4 expression levels were verified by real time RT-PCR (fold change; 24744, 8563, respectively) and Western analysis (B) C. Cell viability assay, D. Invasion assay, E. Wound healing assay, F. Flow cytometric analysis of apoptosis in empty, RECK and Smad4 transfected T24 cells. Data are the mean ± S.D. of four independent experiments. G. beta-catenin expression in nuclear fraction, CREB was used as control.
Table 1.
Primer sequences used for plasmid construction.