Figure 6.

Effects of CWS-loaded formulations on tumor regression in an orthotopic bladder cancer mouse model. (A,B) Luciferase gene-expressing MBT2 (MBT2-Luc) cells were implanted inside the bladder of C3H/He mice. MBT2-Luc tumors in the bladders of C3H mice were detected by IVIS (Perkin Elmer, Waltham, MA, USA). Upon the luminescent intensity being captured via BLS, Veh, CWS-Nano-CL, or CWS-Nano-CL-chitosan were instilled through a catheter into the bladder lumen and remained in situ for 2 h. In all of the statistical analyses, two-sided p-values less than 0.05 were considered statistically significant. ROI time plots for quantitative comparison (Veh/CWS-Nano-CL, Veh/CWS-Nano-CL-chitosan; ** p < 0.005). Data represent the means ± SD (n = 7). (C) CWS-Nano-CL and CWS-Nano-CL-chitosan induced cleaved PARP by activation of AMPK in orthotopic bladder cancer mice. The blots are representative of three independent experiments. All of the mice were sacrificed, and whole bladders of mice were collected from them. Western blot analysis was performed on the bladder tissues of C3H/He mice, and each bladder tumor number is labeled on top o the bands. The blots are representative of three independent experiments. The quantification graphs represent Cl’vd PARP/Actin and p-AMPK/AMPK ratios determined by densitometric analyses. All expression ratios were normalized to the untreated group. Abbreviations: Veh, Vehicle; CWS-Nano-CL, CWS nanoparticles encapsulated with conventional liposomes; CWS-Nano-CL-chitosan, chitosan-coated CWS-Nano-CL; BLS, bioluminescence signal; ROI, region of interest.