title: The natural triterpene maslinic acid induces apoptosis in HT29 colon cancer cells by a JNK-p53-dependent mechanism.
abstract: BACKGROUND
  Maslinic acid, a pentacyclic triterpene found in the protective wax-like coating of the leaves and fruit of Olea europaea L., is a promising agent for the prevention of colon cancer. We have shown elsewhere that maslinic acid inhibits cell proliferation to a significant extent and activates mitochondrial apoptosis in colon cancer cells. In our latest work we have investigated further this compound's apoptotic molecular mechanism.
  
  
  METHODS
  We used HT29 adenocarcinoma cells. Changes genotoxicity were analyzed by single-cell gel electrophoresis (comet assay). The cell cycle was determined by flow cytometry. Finally, changes in protein expression were examined by western blotting. Student's t-test was used for statistical comparison.
  
  
  RESULTS
  HT29 cells treated with maslinic acid showed significant increases in genotoxicity and cell-cycle arrest during the G0/G1 phase after 72 hours' treatment and an apoptotic sub-G0/G1 peak after 96 hours. Nevertheless, the molecular mechanism for this cytotoxic effect of maslinic acid has never been properly explored. We show here that the anti-tumoral activity of maslinic acid might proceed via p53-mediated apoptosis by acting upon the main signaling components that lead to an increase in p53 activity and the induction of the rest of the factors that participate in the apoptotic pathway. We found that in HT29 cells maslinic acid activated the expression of c-Jun NH2-terminal kinase (JNK), thus inducing p53. Treatment of tumor cells with maslinic acid also resulted in an increase in the expression of Bid and Bax, repression of Bcl-2, release of cytochrome-c and an increase in the expression of caspases -9, -3, and -7. Moreover, maslinic acid produced belated caspase-8 activity, thus amplifying the initial mitochondrial apoptotic signaling.
  
  
  CONCLUSIONS
  All these results suggest that maslinic acid induces apoptosis in human HT29 colon-cancer cells through the JNK-Bid-mediated mitochondrial apoptotic pathway via the activation of p53. Thus we propose a plausible sequential molecular mechanism for the expression of the different proteins responsible for the intrinsic mitochondrial apoptotic pathway. Further studies with other cell lines will be needed to confirm the general nature of these findings.
authors.affiliation: Department of Biochemistry and Molecular Biology I, University of Granada, 18071 Granada, Spain.
authors.forename: Fernando J
authors.initials: FJ
authors.lastname: Reyes-Zurita
authors.forename: Gisela
authors.initials: G
authors.lastname: Pachón-Peña
authors.forename: Daneida
authors.initials: D
authors.lastname: Lizárraga
authors.forename: Eva E
authors.initials: EE
authors.lastname: Rufino-Palomares
authors.forename: Marta
authors.initials: M
authors.lastname: Cascante
authors.forename: José A
authors.initials: JA
authors.lastname: Lupiáñez
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D051023:BH3 Interacting Domain Death Agonist Protein / Q000819:agonists; D020169:Caspases / Q000378:metabolism; D003110:Colonic Neoplasms / Q000473:pathology; D020552:Comet Assay; D045304:Cytochromes c / Q000378:metabolism; D019073:HT29 Cells; D006801:Humans; D048670:MAP Kinase Kinase 4 / Q000378:metabolism*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000037:antagonists & inhibitors; D014315:Triterpenes / Q000494:pharmacology*; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism*; D051028:bcl-2-Associated X Protein / Q000819:agonists; D009828:Oleanolic Acid / Q000031:analogs & derivatives
chemical_list: D000970:Antineoplastic Agents; D051023:BH3 Interacting Domain Death Agonist Protein; D020169:Caspases; D045304:Cytochromes c; D048670:MAP Kinase Kinase 4; D019253:Proto-Oncogene Proteins c-bcl-2; D014315:Triterpenes; D016159:Tumor Suppressor Protein p53; D051028:bcl-2-Associated X Protein; C412811:maslinic acid; D009828:Oleanolic Acid