title: CDODA-Me decreases specificity protein transcription factors and induces apoptosis in bladder cancer cells through induction of reactive oxygen species.
abstract: The objective is to determine whether methyl 2-cyano-3,11-dioxo-18b-olean-1,12-dien-30-oate (CDODA-Me) has therapeutic potential in bladder cancer. We investigated the effects of CDODA-Me on the growth and survival of bladder cancer cells, and expression of specificity protein (Sp) transcription factors that regulate genes associated with cancer cell proliferation and survival.
  
  
  
  J82, RT4P, and 253JB-V bladder cancer cell lines were treated with vehicle alone or with CDODA-Me with or without the antioxidant l-glutathione. Cell viability and DNA fragmentation were measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and propidium iodide-fluorescence-activated cell sorting (FACS) analysis, respectively. Intracellular reactive oxygen species (ROS) were measured by 2',7'-dichlorofluorescin diacetate-FACS analysis. We assessed CDODA's effects on the levels of Sp and Sp-regulated proteins and induction of apoptosis in bladder cancer cells by Western blotting. We also assessed the anticancer effects of CDODA-Me in nude mice bearing RT4v6 bladder cancer.
  
  
  
  3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and FACS analysis revealed that CDODA-Me inhibited the proliferation and survival of the 3 bladder cancer cell lines in a dose-dependent manner. FACS analysis also indicated that CDODA-Me-induced intracellular ROS, and Western blot analysis indicated that CDODA-Me decreased levels of Sp and Sp-regulated proteins and induced apoptosis in a dose-dependent and time-dependent manner. l-Glutathione attenuated CDODA-Me's down-regulation of Sp and Sp-regulated proteins. Compared with the control treatment, CDODA-Me substantially inhibited tumor growth in vivo.
  
  
  
  CDODA-Me has antineoplastic activity in bladder cancer cells by inducing ROS, which down-regulate Sp and Sp-regulated proteins. Thus, CDODA-Me has therapeutic potential in bladder cancer, and additional studies of the agent's efficacy and mode of action are warranted.
authors.affiliation: Department of Urology, The University of Texas MD Anderson Cancer Center, Houston, TX.
authors.forename: Hisashi
authors.initials: H
authors.lastname: Takeuchi
authors.affiliation: Department of Urology, The University of Texas MD Anderson Cancer Center, Houston, TX.
authors.forename: Rikiya
authors.initials: R
authors.lastname: Taoka
authors.affiliation: Department of Urology, The University of Texas MD Anderson Cancer Center, Houston, TX.
authors.forename: Chinedu O
authors.initials: CO
authors.lastname: Mmeje
authors.affiliation: Department of Urology, The University of Texas MD Anderson Cancer Center, Houston, TX.
authors.forename: Goodwin G
authors.initials: GG
authors.lastname: Jinesh
authors.affiliation: Veterinary Physiology and Pharmacology, Texas A&M University, College Station, TX.
authors.forename: Stephen
authors.initials: S
authors.lastname: Safe
authors.affiliation: Department of Urology, The University of Texas MD Anderson Cancer Center, Houston, TX. Electronic address: akamat@mdanderson.org.
authors.forename: Ashish M
authors.initials: AM
authors.lastname: Kamat
mesh_terms: D000818:Animals; D017209:Apoptosis; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D006034:Glycyrrhetinic Acid / Q000031:analogs & derivatives* / Q000494:pharmacology; D006801:Humans; D008297:Male; D051379:Mice; D008819:Mice, Nude; D017382:Reactive Oxygen Species / Q000378:metabolism*; D051703:Sp Transcription Factors / Q000378:metabolism*; D001749:Urinary Bladder Neoplasms / Q000188:drug therapy*; D023041:Xenograft Model Antitumor Assays
chemical_list: D017382:Reactive Oxygen Species; D051703:Sp Transcription Factors; C543611:methyl 2-cyano-3,11-dioxo-18beta-olean-1,12-dien-30-oate; D006034:Glycyrrhetinic Acid
keywords: Anticancer agents; Antioxidants; Bladder cancer; Reactive oxygen species (ROS); Specificity protein (Sp) transcription factors; Targeted therapy