title: Methylferulate from Tamarix aucheriana inhibits growth and enhances chemosensitivity of human colorectal cancer cells: possible mechanism of action.
abstract: BACKGROUND
  Natural products are valuable sources for anticancer agents. In the present study, methylferulate (MF) was identified for the first time from Tamarix aucheriana. Spectral data were used for identification of MF. The potential of MF to control cell growth, cell cycle, apoptosis, generation of reactive oxygen species (ROS), cancer cell invasion, nuclear factor kappa B (NFkB) DNA-binding activity and proteasomal activities, as well as the enhancement of chemosensitivity in human colorectal cancer cells, were evaluated. The possible molecular mechanism of MF's therapeutic efficacy was also assessed.
  
  
  METHODS
  Column chromatography and spectral data were used for isolation and identification of MF. MTT, immunofluorescence, flow cytometry, in vitro invasion, fluoremetry, EIA and Real time qPCR were used to measure antiproliferative, chemo-sensitizing effects and other biochemical parameters.
  
  
  RESULTS
  MF showed a dose-dependent anti-proliferative effect on colorectal cancer cells (IC50= 1.73 - 1.9 mM) with a nonsignificant cytotoxicity toward normal human fibroblast. Colony formation inhibition (P ≤ 0.001, 0.0001) confirmed the growth inhibition by MF. MF arrested cell cycle progression in the S and G2/M phases; induced apoptosis and ROS generation; and inhibited NF-kB DNA-binding activity, proteasomal activities and cell invasion in colorectal cancer cells. MF up-regulated cyclin-dependent kinase inhibitors (p19INK4D, p21WAF1/CIP1, p27KIP1), pro-apoptotic gene expression (Bax, Bad, Apaf1, Bid, Bim, Smac) and caspases (caspase 2, 3, 6, 7, 8, 9). Moreover, MF down-regulated cyclin-dependent kinases (Cdk1, Cdk2) and anti-apoptotic gene expression (c-IAP-1, c-IAP-2, Bcl2,FLIP). In addition, MF differentially potentiated the sensitivity of colorectal cancer cells to standard chemotherapeutic drugs.
  
  
  CONCLUSIONS
  MF showed a multifaceted anti-proliferative and chemosensitizing effects. These results suggest the chemotherapeutic and co-adjuvant potential of MF.
authors.affiliation: Molecular Biology Program, Department of Biological Sciences, Faculty of Science, P. O. Box 5969, Safat, 13060, Kuwait. abazams@yahoo.com.
authors.forename: Mohamed Salah I
authors.initials: MS
authors.lastname: Abaza
authors.affiliation: Biochemistry Program, Department of Biological Sciences, Faculty of Science, P. O. Box 5969, Safat, 13060, Kuwait.
authors.forename: Mohammad
authors.initials: M
authors.lastname: Afzal
authors.affiliation: Microbiology and Immunology Department, Faculty of Medicine, Kuwait University, Kuwait City, Kuwait.
authors.forename: Raja'a J
authors.initials: RJ
authors.lastname: Al-Attiyah
authors.affiliation: Molecular Biology Program, Department of Biological Sciences, Faculty of Science, P. O. Box 5969, Safat, 13060, Kuwait.
authors.forename: Radhika
authors.initials: R
authors.lastname: Guleri
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D002109:Caffeic Acids / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects*; D015179:Colorectal Neoplasms; D006801:Humans; D010936:Plant Extracts / Q000737:chemistry*; D032363:Tamaricaceae / Q000737:chemistry*
chemical_list: D000970:Antineoplastic Agents; D002109:Caffeic Acids; D010936:Plant Extracts; C530806:methyl ferulate
keywords: Apoptosis; Chemosensitization; Colorectal cancer; Methylferulate; Molecular mechanisms; Tamarix aucheriana