Fig. 7. The inhibitory effect of BMS-777607 on RON-mediated activity and the suppressive effect of ectopic expression of miR-659-3p on bladder cancer cells tumorigenic in vivo.

A J82 cells were infected with a lentivirus package to overexpress RON (J82-RON). An empty lentivirus vector package was used as a negative control to generate J82-NC cells. These two cell lines were inoculated into thymic nude mice, respectively, and tumor growth was monitored daily. Upon the tumor volume reaching ~100 mm³ (~14 days), mice were divided into 2 groups. Mice in the experimental group (J82-RON, 5 mice per group) were treated with BMS-777607 at 25 mg/kg/day. The mice in the control group (J82-RON, 5 mice per group) and mice from the J82-NC group, 5 mice per group) were administered with solvents. Images of tumors from individual mice were captured on day 32 following the injection of J82 cells. B The tumor growth was assessed daily and represented in a line chart. The weight of the tumor is shown in (C). D Upon the end of the study, all animals were dissected to confirm any organ metastases. Images of hepatic tissues were presented. E Hematoxylin and eosin (H&E) staining was performed to for pathological examination to find any metastatic tumors (Scale bar 500 nm). F T24 cells were transfected with LV-miR-659-3p to create T24-659-3p cells, which were subsequently transfected with lentivirus overexpressing RON to generate T24-659-3p-RON cells. The resulting T24 cells were then subcutaneously injected into the armpits of nude mice. Images of tumors from each group were captured on day 32 following the injection of transfected T24 cells. G The tumor growth was assessed periodically and represented in a line chart. The final weight of the tumor was shown in (H). I Then, all animals were further dissected to observe organ metastasis, and images of livers were illustrated. J Hematoxylin and eosin (H&E) staining was performed to detect metastatic livers. (Scale bar 500 nm).