#B193 Pro-Apoptotic Properties of Novel Celecoxib-Derivatives in Bladder Cancer Cell Lines. Peter S. Carlton,1 Steven K. Clinton,1 Ching-Shih Chen,2 Dahlys R. Hoot,1 Valerie L. DeGroff,1 Robert R. Bahnson.3 The Ohio State University, College of Medicine and Public Health, Department of Internal Medicine, Division of Hematology and Oncology,1 Columbus, Ohio, The Ohio State University, College of Pharmacy, Division of Medicinal Chemistry and Pharmacognosy,2 Columbus, Ohio, The Ohio State University, College of Medicine and Public Health, Department of Surgery, Division of Urology,3 Columbus, Ohio. Bladder cancer is the 6th most commonly diagnosed cancer in the United States, and is predicted to cause 12,500 deaths in 2003. Clearly, innovative and effective approaches for the prevention and treatment of bladder cancer are needed. Epidemiologic and animal model studies provide significant rationale for the development of non-steroidal anti-inflammatory drugs (NSAIDs) as chemopreventive agents in the bladder. NSAIDs classified as selective cyclooxygenase (COX)-2 inhibitors, including celecoxib, are particularly appealing due to their limited toxicity and proven ability to inhibit bladder cancer in rodents. Using celecoxib as a molecular starting point, we applied computer modeling and structure-activity analysis to design novel derivative compounds with potentially greater chemopreventive activity. Initially, a series of celecoxib derivatives (OSU-02062, OSU-02064 and OSU-02074) was screened for cytotoxicity in HT1376, J82, RT4, T24, TCCSUP and UMUC3 bladder cancer cells using a MTS cell viability assay. Our results indicate that J82, RT4, T24 and TCCSUP cells were more sensitive to celecoxib-derivative treatment compared to HT1376 and UMUC3 cells, and that OSU-02074 was the most cytotoxic derivative in each cell line. In particular, OSU-02074 at 5 ��M inhibited RT4 and J82 cell viability by 70% and 90%, respectively. While OSU-02062 and OSU-02064 also significantly inhibited cell viability, effects of the parent compound, celecoxib (1 − 20 ��M), were negligible in all of the cell lines tested. We also conducted assays to determine if the observed cytotoxicity of celecoxib-derivatives in bladder cancer cells was related to the induction of apoptosis. In RT4 and J82 cells treated with OSU-02074 (5 ��M), we observed apoptosis-related DNA condensation and fragmentation by 4��-6-diamidino-2-phenylindole (DAPI) staining. The pro-apoptotic activity of OSU-02074 in RT4 and J82 cells was confirmed by annexin V binding and caspase-3/7 activity assays, and by Western blot for cleaved poly-(ADP-ribose) polymerase (PARP). Our data indicate enhanced cytotoxicity of certain celecoxib-derivatives compared to the parent compound and suggest that this effect is related, in part, to the induction of apoptosis. In addition, we have preliminary data to suggest that celecoxib-derivative-induced apoptosis in J82 cells is associated with de-phosphorylation of the anti-apoptotic protein, Akt. Ongoing studies will elucidate additional mechanisms whereby these agents modulate apoptotic-signaling pathways. Together, our studies support further pre-clinical development of celecoxib-derivatives as agents for the prevention or treatment of bladder cancer. Frontiers in Cancer Prevention Research, 2003 AACR Ann's NOTE: In discussions we found that these products will be moving to NCI for testing in human experimental subjects. (Clinical trials) Remember we are NOT Doctors and have NO medical training. This site is like an Encylopedia - there are many pages, many links on many topics. Support our work with any size DONATION - see left side of any page - for how to donate. You can help raise awareness of CAM.