title: The position of the amino group on the benzene ring is critical for mesalamine's improvement of replication fidelity.
abstract: BACKGROUND
  Individuals with ulcerative colitis are at high risk of developing colitis-associated cancer. 5-Aminosalicylate (5-ASA) protects from cancer by its antiinflammatory activity as well as by altering cell growth, inducing apoptosis, and reducing replication errors. So far neither 5-ASA's structural specificity nor its pharmacophore group have been identified. Here we compared 5-ASA with its analogs (4-ASA and 3-ASA) and its metabolite N-acetyl-5-ASA (NAc-5-ASA).
  
  
  METHODS
  Superoxide scavenging was analyzed by lucigenin-amplified chemiluminescence. Cell growth, cell cycle distribution, and replication fidelity at a (CA)13 microsatellite were measured in HCT116 and HT29 colon epithelial cells by MTT and flow cytometry. Nuclear protein extracts were blotted for replication protein A (RPA), claspin, p53, and p53(Ser15).
  
  
  RESULTS
  All compounds inhibited the growth of colon epithelial cells at a similar level and displayed potent scavenging properties, with 3-ASA being the most active, followed by 5-ASA, 4-ASA, and NAc-5-ASA. Besides 5-ASA, only 4-ASA caused an increase in the S-phase population (56%-69% and 49%-62% in HCT116 and HT29 cells, respectively). This was accompanied by nuclear recruitment of replication proteins RPA and claspin as well as phosphorylation of p53(Ser15), both of which were weaker or absent with 3-ASA or NAc-5-ASA. 5-ASA was the only compound that lowered mutations at a (CA)13 microsatellite.
  
  
  CONCLUSIONS
  5-ASA shares its growth inhibitory and superoxide scavenging properties with its structural analogs and metabolite, but the position of the amino group is critical for reducing replication errors.
authors.affiliation: Medical University of Vienna, Department of Internal Medicine III, Division of Gastroenterology and Hepatology, and Christian Doppler Laboratory on Molecular Cancer Chemoprevention, Vienna, Austria.
authors.forename: Christoph
authors.initials: C
authors.lastname: Campregher
authors.forename: Maria Gloria
authors.initials: MG
authors.lastname: Luciani
authors.forename: Peter
authors.initials: P
authors.lastname: Biesenbach
authors.forename: Rayko
authors.initials: R
authors.lastname: Evstatiev
authors.forename: Alex
authors.initials: A
authors.lastname: Lyakhovich
authors.forename: Christoph
authors.initials: C
authors.lastname: Gasche
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000494:pharmacology*; D001554:Benzene / Q000737:chemistry*; D015153:Blotting, Western; D002453:Cell Cycle / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D004261:DNA Replication / Q000187:drug effects*; D005434:Flow Cytometry; D019073:HT29 Cells / Q000187:drug effects; D006801:Humans; D019804:Mesalamine / Q000494:pharmacology*; D018895:Microsatellite Repeats; D009504:Neutrophils / Q000378:metabolism; D051716:Replication Protein A / Q000378:metabolism*; D013481:Superoxides / Q000378:metabolism; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D000894:Anti-Inflammatory Agents, Non-Steroidal; C488158:CLSPN protein, human; D051716:Replication Protein A; C495901:TP53 protein, human; D016159:Tumor Suppressor Protein p53; D013481:Superoxides; D019804:Mesalamine; D001554:Benzene