Colon26 NL 1-7 mouse colon carcinoma cells were cultured in DMEM/Ham F12 medium supplemented with 10 % FBS at 37 C in a CO2 incubator. HUVECs were seed o-n gelatin coated 35mm dishes at 105 cells/dish and incubated over night. After changing of culture medium to endothelial basal medium 2 supplemented with 0. Five hundred fetal bovine contact us serum, the cells were treated with free SU1498 dissolved in DMSO, PEG modified liposomal SU1498, and APRPGPEG modified liposomal SU1498 at 1_M of the ultimate concentration of SU1498 for 3 h. Then, recombinanthumanVEGF165 was put into the cells, and the cells were incubated for another 48 h. Colon26 NL 1-7 cellswere seeded, and the cellswere incubated overnight in DMEM/Ham F12medium supplemented with 10% FBS at 37 C. Then, the cells were further incubated for 48 h and treated with the examples. Finally, the viable cells were stained with crystal violet, and the dye was extracted with 33-in acetic acid and measured at absorbance of 570 nm as described previously. Colon26 NL 1-7 cells were implanted subcutaneously into the rear flank of 5 week-old BALB/c male mice. From days 3 to 11 after cyst implantation, each test, specifically, PEG APRPG PEG Lip SU1498, Lip SU1498, and 0. Every other day 3m sucrose solution, was injected intravenously. O-n day 13, the mice were sacrificed Urogenital pelvic malignancy under anesthesia with diethyl ether, and the tumors were excised. The tumefaction cells were frozen at?80 C and mounted on OCT compound. The tumor tissue sections were prepared with microtome and mounted onto Matsunami adhesive silane coated slide glass. Immunohistochemical staining against CD31 was conducted described previously with some modifications. The parts were fixed with ice-cold acetone, cleaned with phosphate buffered saline, and blocked endogenous peroxidase activity with three or four H2O2 in PBS. Low certain protein bindings were blocked with 1000 bovine serum albumin dissolved in PBS. Then, a murine anti CD31 monoclonal antibody was put into the sections and extra staining was done with VECTASTAIN ABC system according to the manufacturers guidelines. These sections were rinsed and counterstained with Mayers hematoxylin. For quantification contact us of cyst blood vessels, three of high vessel density places per area were selected and captured using Olympus IX71. CD31 positive spot was quantified with ImageJ computer software. Colon26 NL 1-7 showing micewere prepared as described above. Each liposomal SU1498 o-r 0. 3M sucrose solution was applied by the following two different schedules; intravenously injected from days 3 to 1-1 every other day after tumor implantation; intraperitoneally injected from days 1 to 12 every day after tumor implantation. On tumor in vivo because SU1498 is nearly insoluble in water, we could not analyze the effect of the free drug.