title: Synergism of Fas-mediated apoptosis and tumor necrosis factor on adriamycin cytotoxicity to transitional cell carcinoma.
abstract: OBJECTIVE
  To explore the accurate Fas antigenic expression in transitional cell carcinoma (TCC) cells and and compare its synergistic modulation on adriamycin cytotoxicity with alpha-tumor necrosis factor (TNF-alpha).
  
  
  METHODS
  The expression of Fas antigen in normal urothelium and TCC cell lines was measured by flow cytometry. The Fas/Fas ligand reaction-induced cytotoxic changes in tumor cells were evaluated by microculture MTT technique. The synergism efficacy of Fas-mediated apoptosis and TNF-alpha on adriamycin cytotoxicity of TCC cells was analyzed. The Fas/Fas ligand-induced apoptosis in tumor cells was studied by annexin-V apoptotic cell expression and DNA ladder fragmentation analysis.
  
  
  RESULTS
  75% of TCC cell lines showed Fas antigen expression. The Fas expression was not influenced by in vitro growth density of tumor cells. Apoptosis of TCC cells was observed during 48 h of treatment after activation of the Fas/Fas ligand pathway. TNF-alpha had a higher cytotoxicity activity on TCC cells than Fas ligand (17 vs. 0.7%) while combination of both reagents had 30% increased synergistic cytotoxicity. The increasing rate of apoptotic body after 3 days of treatment was 74% in adriamycin, 32% in Fas ligand, 60% in TNF-alpha, 69% in Fas and adriamycin combination and Fas and TNF-alpha combination, 103% in combination of TNF-alpha and adriamycin, and 136% in combination of these three reagents individually. In the DNA ladder analysis, Fas ligand had a 1.5-fold increase of DNA fragmentation when compared with adriamycin while TNF-alpha had a 4.4-fold increase and triple combination had a 5.5-fold increase after 3 days of treatment.
  
  
  CONCLUSIONS
  Although the Fas antigen expression is common in TCC cells and apoptosis can be activated by the Fas/Fas ligand pathway activation, the synergistic cytotoxicity of adriamycin by the Fas/Fas ligand pathway is lower than that of TNF-alpha.
authors.affiliation: Uro-Oncology Laboratory, Division of Urology, Department of Surgery, Tri-Service General Hospital, No. 325, Sec. 2 Cheng-Gung Road, Nei-Hu, Taipei 114, Taiwan/ROC. yuds@ms21.hinet.net
authors.forename: Dah-Shyong
authors.initials: DS
authors.lastname: Yu
authors.forename: Dar-Shih
authors.initials: DS
authors.lastname: Hsieh
authors.forename: Hong-I
authors.initials: HI
authors.lastname: Chen
authors.forename: Sun-Yran
authors.initials: SY
authors.lastname: Chang
mesh_terms: D017209:Apoptosis; D002295:Carcinoma, Transitional Cell / Q000473:pathology*; D004317:Doxorubicin / Q000633:toxicity*; D053222:Fas Ligand Protein; D006801:Humans; D008562:Membrane Glycoproteins / Q000494:pharmacology*; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*; D001749:Urinary Bladder Neoplasms / Q000473:pathology*; D019014:fas Receptor / Q000494:pharmacology*
chemical_list: C505661:FASLG protein, human; D053222:Fas Ligand Protein; D008562:Membrane Glycoproteins; D014409:Tumor Necrosis Factor-alpha; D019014:fas Receptor; D004317:Doxorubicin