Research Paper Volume 11, Issue 24 pp 12624—12640

Long noncoding RNA NNT-AS1 enhances the malignant phenotype of bladder cancer by acting as a competing endogenous RNA on microRNA-496 thereby increasing HMGB1 expression

Figure 5. MiR-496 performs its tumor-suppressive actions in bladder cancer cells by decreasing HMGB1 expression. (A) The miR-496 mimics in combination with either the HMGB1-overexpressing plasmid pc-HMGB1 or the empty pcDNA3.1 vector was cotransfected into T24 and TCC-SUP cells. At 72 h post-transfection, western blotting was performed to analyze HMGB1 expression. *P < 0.05 vs. the miR-NC group. #P < 0.05 vs. the miR-496 mimics+pcDNA3.1 group. (B, C) The proliferative and apoptotic activities of T24 and TCC-SUP cells after cotransfection with the miR-496 mimics and either pc-HMGB1 or pcDNA3.1 were evaluated through the CCK-8 assay and flow-cytometric analysis, respectively. *P < 0.05 vs. the miR-NC group. #P < 0.05 vs. group miR-496 mimics+pcDNA3.1. (D, E) Transwell migration and invasion assays were conducted to examine the migratory and invasive abilities of T24 and TCC-SUP cells after cotransfection with the miR-496 mimics and either pc-HMGB1 or pcDNA3.1. *P < 0.05 vs. group miR-NC. #P < 0.05 vs. the miR-496 mimics+pcDNA3.1 group.