Briefly, SW620 cells were transiently transfected MEK162 ARRY-438162 with scramble siRNA, and human xIAP and cIAP1 specific siRNAs, respectively, using Lipofectamine 2000 for approximately Inhibitors,Modulators,Libraries 24 h. Cells were then harvested. Part of the cells were used for RT PCR analysis of xIAP and cIAP expression. Another part of the cells were cultured in the absence or presence of FasL for approximately 24 h and then analyzed for apoptosis. Liver toxicity analysis LCL85 was injected to BALB/c mice i. v. Peripheral blood was collected from mice 3 days later using Multivette 600 Z gel tubes. Inhibitors,Modulators,Libraries Serum was separated by centrifugation and measured for complete liver enzyme profile at Georgia Laboratory Animal Diagnostic Service. Colon cancer experimental lung metastasis Colon 26 cells were injected to BALB/c mice iv.

LCL85 was injected iv to tumor bearing mice at days 3, 6, 9 and 12 after tumor injection. Mice were sacrificed at day 14 and analyzed for lung metastasis as previously described. Breast cancer spontaneous lung metastasis 4 T1 cells were injected to the mammary fat pad. LCL85 was injected to the tumor Inhibitors,Modulators,Libraries bearing mice at days 7, 10, 13, and 16 after tumor injection. Mice were sacrificed 29 days after tumor injection, and analyzed for primary tumor growth and lung metastasis. To determine the efficacy of LCL85 on metastasis, 4 T1 cells were injected to the mammary fat pad. Primary tumors were surgically removed 16 days later. Mice were treated with LCL85 at days 10, 13, and 16 after surgery. Mice were sacrificed and analyzed for lung metastasis 19 days after surgery.

Statistical analysis Where indicated, data were represented Inhibitors,Modulators,Libraries as the mean SD. Statistical analysis was performed using two sided t test, with p values 0. 05 considered statistically significant. Results Ceramide analog effectively sensitizes metastatic human colon and breast cancer cell apoptosis resistance Ceramide analogs of B13 and LCL85 were tested for their cytotoxicity against human colon carcinoma cell lines. Cell growth inhibition assays indicated that B13 and LCL85 are both cytotoxic at high doses. LCL85 represents a unique compound since it is highly cytotoxic at high doses, but exhibited almost no cytoto xicity at low doses. Because our objective was to test the hypothesis that ceramide analogs are effective apoptosis sensitizers for Fas Inhibitors,Modulators,Libraries mediated apoptosis in human colon carcinoma cells, we chose LCL85 for this study.

Next, eleven human colon carcinoma cell lines were cul tured in the presence of a sublethal dose of LCL85 and various doses of FasL, and analyzed for tumor cell viability. Four of the 6 primary colon carcinoma cell lines are highly sensitive to FasL induced apoptosis, and LCL85 exhibited minimal or no sensitization effects on these 4 sensitive cell selleckchem Dasatinib lines. On the other hand, the other 2 primary human colon carcinoma cell lines RKO and SW116 are resistant to Fas mediated apoptosis.