Metastatic model

The hemispleen technique of tumor inoculation was performed on day 0. In brief, after anesthetizing the mouse, a left subcostal incision was made, and the spleen was eviscerated, clipped, and hemisected. One half of the spleen was injected with 2 × 10⁵ KPC cells resuspended in 100 μl PBS and flushed with 150 μl PBS in the same syringe. Cells were injected slowly into the exposed hemispleen, while the syringe was kept upright at all times to ensure the 150-μl PBS flush remained as free from tumor cells as possible. The splenic vessels were then clipped, and the injected hemispleen was resected to remove residual tumor cells. Following this procedure, diffuse liver metastases develop, and we previously reported that all untreated mice die in 4–6 wk. Mice were randomized to each treatment group after the surgery.