Figure 6. Bladders of female C57/BL6 mice were orthotopically implanted with MB49 cells (1.2×105) and divided into 4 groups randomly (n = 12 each group).

Orthotopic bladder cancer implantation was processed for mice in groups I, II, III, and IV. Group V was mice without treatment without cancer cell implantation. Groups I and II were treated with either PBS or 800mg/kg/d metformin in PBS orally. Oral treatment started at day 2 and treated daily. Groups III and IV were intravesically treated with 50 μl either 80mM or 320mM metformin, respectively in PBS per mouse, starting at day 2, twice per week for two weeks. A. Kaplan-Meier survival analysis of five groups. Death of mice was checked daily and cumulative survival rate was plotted against the time course. Life span was 12.5 days vs 16 days in groups I and II mice, respectively. The weight of mice was measured daily B. and all living mice were sacrificed at day 28. C. Weights of mouse bladders including those died before the end of experiment were measured. All bladder tissues were collected and fixed. Histological sections from these tissues were subjected to H&E stain or immunohistochemistry for Ki67 to confirm the presence or absence of tumors D. & E.