The drug even appeared to have a postantibiotic effect in infected mice as seen

Treatment of Established Human Colon Carcinomas Growing in the Cecum of Athymic Nude Mice Fourteen days after injection of tumor cells. Irinotecan was kept at room temperature and dissolved in 0. 9% NaCl on the day of intraperitoneal injection. Celecoxib Primary antibodies used were as follows: rabbit anti phosphorylated EGFR, mouse anti EGFR, mouse anti TGF rabbit anti EGF, rat anti mouse CD31, and rabbit anti Ki 67 antigen for immunohistochemistry, and rabbit anti EGFR forWestern blot analysis. The following secondary antibodies were used for colorimetric immunohistochemistry: peroxidase conjugated goat anti rabbit IgG, peroxidase conjugated goat anti mouse IgG, and peroxidase conjugated goat anti rat IgG. The following fluorescent Eumycetoma secondary antibodies were used: Cy3 conjugated goat anti rabbit IgG, Cy3 conjugated goat anti mouse IgG, Cy3 conjugated goat anti rat IgG, and Cy5 conjugated goat anti rat IgG. The following secondary antibodies were used for Western blot analysis: peroxidase conjugated goat anti rabbit IgG. Terminal deoxynucleotidyl transferase mediated nick end labeling staining was done using a commercial apoptosis detection kit with modifications. Animals and Orthotopic Implantation of Tumor Cells Male athymic nude mice were purchased from the Animal Production Area of the National Cancer Institute Frederick Cancer Research and Development Center. The mice were housed and maintained under specific pathogen free conditions in facilities approved by the American Association for Accreditation of Laboratory Animal Care and in accordance with current regulations and standards of the US Department of Agriculture, the US Department of Health and Human Services, and the National Institutes of Health. The mice were used in accordance with institutional guidelines when they were 8 to 12 weeks old. To produce cecal tumors, SW620CE2 WT, SW620CE2 nontargeting shRNA, and SW620CE2 TGF shRNA cells were harvested from subconfluent cultures by a brief exposure to 0. 25% trypsin and 0. 02% EDTA. BAY 11-7082 Trypsinization was stopped with medium containing 10% fetal bovine serum, and the cells were then washed once in serum free medium and resuspended in Hanks balanced salt solution. Only suspensions consisting of single cells with 90% viability were used. A total of 5 105 cells in 50 ul of Hanks balanced salt solution were injected into the cecal wall of nude mice under a dissecting microscope as described previously. when cecal tumors reached the size of 4 to 5 mm in diameter, groups of 10 mice each were randomly assigned to receive one of the following four treatments: 1) oral administration of water diluted at 1:20 with DMSO 0. 5% Tween 80 three times per week and i. p. injection of PBS once a week, 2) oral administration of PKI166 three times per week and i. p. injection of PBS once a week, 3) oral administration of diluent by three times per week and i.