Expression data from mice bladder with MB49-induced cancer treated with alpha1-oleate alone or in combination Epirubicin or Mitomycin C, short-term effects

Bladder cancer is common and one of the most costly cancer forms, due to a lack of curative therapies. Recently, clinical safety and efficacy of the alpha1-oleate complex was demonstrated in a placebo-controlled study of non-muscle invasive bladder cancer. This study investigated if long-term therapeutic efficacy is improved by repeated treatment cycles and by combining alpha1-oleate with low-dose chemotherapy. Rapidly growing bladder tumors were treated by intravesical instillation of alpha1-oleate, Epirubicin or Mitomycin C alone or in combination. One treatment cycle arrested tumor growth, with a protective effect lasting at least four weeks in mice receiving 8.5 mM of alpha1-oleate alone or 1.7 mM of alpha-oleate combined with Epirubicin or Mitomycin C. Repeated treatment cycles extended protection, defined by a lack of bladder pathology and a virtual absence of bladder cancer-specific gene expression. Synergy with Epirubicin was detected at the lower alpha1-oleate concentration and alpha1-oleate was shown to enhance the uptake and nuclear translocation of Epirubicin, by tumor cells. Effects at the chromatin level affecting cell proliferation were further suggested by reduced BrdU incorporation. In addition, alpha1-oleate triggered DNA fragmentation, defined by the TUNEL assay. The results suggest that bladder cancer development may be prevented long-term in the murine model, by alpha1-oleate alone or in combination with low-dose Epirubicin. In addition, the combination of alpha1-oleate and Epirubicin reduced the size of established tumors. Exploring these potent preventive and therapeutic effects will be of immediate interest in patients with bladder cancer. Bladder cancer was induced in C57BL/6J female mice by intravesical instillation of MB49 (RRID:CVCL7076) mouse bladder carcinoma cells (2 × 10^5 cells in 50 μl cell media) in empty bladders preconditioned by intravesical instillation of 100 μl of poly-L-lysine solution (0.1 mg/ml, 30 minutes). Mice were randomly assigned to the different treatment groups and subjected to 100 μl instillations with alpha1-oleate (1.7 mM or 8.5 mM) or chemotherapeutic drugs (MMC 25 μg, Epirubicin 25 μg), alone or in combination on days 3, 5, 7, 9 and 11. Sham treated mice received PBS and healthy mice bladders were used as control. The catheter was left in place for about one minute and the time to voiding was 2 - 3 hours. Bladder samples were collected on day 12 day or day 28 days (follow-up, 4 weeks). Isolated RNA was subjected to Affymetrix whole genome transcriptomic analysis.