After incubation, cells were trypsi nized and suspended in 1 ml of low serum medium. From each cell suspension 2105 cell were centrifuged and re suspended in 500 ul 1X binding buffer. In each sample 5 ul of Annexin V FITC and 5 ul of PI was added, incubated for 5 minutes selleck bio in dark and immediately analyzed by flow cytometry. The annexin V FITC positive and PI negative cells were considered Inhibitors,Modulators,Libraries to be early apoptotic cells. Animal Studies All animal studies were performed in accordance with the Institutional Animal Care and Use Committee at the University of Texas Southwestern Medical Center. In vivo animal studies were performed using 4 6 weeks old SCID mice purchased from an on campus facility. Tumor growth experiments were per formed by orthotopically Inhibitors,Modulators,Libraries injecting 1106 Panc 1 cells.

Inhibitors,Modulators,Libraries Animals Inhibitors,Modulators,Libraries were examined by ultrasound and randomized into treatment groups when tumor size averaged 500 mm3 at approximately 8 weeks after tumor cell injection. Animals were injected intraperitoneally with PBS, JP and Gem, either alone or in combination. Three mice from each treatment group were sacrificed at 24 hours of therapy while the remaining animals in each group received 2 weeks of therapy prior to sacrifice. Animal survival studies were conducted in an intra peritoneal PDAC tumor model as previously described. Female SCID mice received 0. 75106 human AsPC 1 cells intraperitoneally. The animals were ran domly grouped and treated intra peritoneally with PBS, JP, Gem and DT, either alone or in combination for 14 days or as maintenance therapy.

Animal weight was measured twice weekly and all animals were examined daily for signs of distress or development of jaundice. Moribund mice at risk for distress were euthanized Inhibitors,Modulators,Libraries in accordance with the local animal care committee protocol. Subse quently, animals were examined for presence and extent of intra abdominal tumor. Statistical analysis In vitro cell proliferation assay results are expressed as meanstandard deviation. Statistical significance was analyzed by the two tailed Students t test using Graph Pad Prism 4 Software. For in vivo studies, statistical analysis was per formed by ANOVA for multiple group comparison and Students t test for the individual group comparison. In survival studies, statistical differences were analyzed by nonparametric survival statistics and logrank test. Values of p 0. 05 were considered to represent statistically sig nificant group difference. Results Effect of JP and Gem on PDAC cell proliferation In vitro WST 1 assay selleck chemicals Imatinib was performed to examine the effect of JP and Gem on PDAC cell proliferation. In AsPC 1 cells, JP and Gem significantly inhibited the cell proliferation. After 72 hours of incubation, JP and Gem inhibited the AsPC 1 cell prolifera tion by 31% and 58%, respectively.