title: Antimutagenicity of cinnamaldehyde and vanillin in human cells: Global gene expression and possible role of DNA damage and repair.
abstract: Vanillin (VAN) and cinnamaldehyde (CIN) are dietary flavorings that exhibit antimutagenic activity against mutagen-induced and spontaneous mutations in bacteria. Although these compounds were antimutagenic against chromosomal mutations in mammalian cells, they have not been studied for antimutagenesis against spontaneous gene mutations in mammalian cells. Thus, we initiated studies with VAN and CIN in human mismatch repair-deficient (hMLH1(-)) HCT116 colon cancer cells, which exhibit high spontaneous mutation rates (mutations/cell/generation) at the HPRT locus, permitting analysis of antimutagenic effects of agents against spontaneous mutation. Long-term (1-3 weeks) treatment of HCT116 cells with VAN at minimally toxic concentrations (0.5-2.5mM) reduced the spontaneous HPRT mutant fraction (MF, mutants/10(6) survivors) in a concentration-related manner by 19-73%. A similar treatment with CIN at 2.5-7.5microM yielded a 13-56% reduction of the spontaneous MF. Short-term (4-h) treatments also reduced the spontaneous MF by 64% (VAN) and 31% (CIN). To investigate the mechanisms of antimutagenesis, we evaluated the ability of VAN and CIN to induce DNA damage (comet assay) and to alter global gene expression (Affymetrix GeneChip) after 4-h treatments. Both VAN and CIN induced DNA damage in both mismatch repair-proficient (HCT116+chr3) and deficient (HCT116) cells at concentrations that were antimutagenic in HCT116 cells. There were 64 genes whose expression was changed similarly by both VAN and CIN; these included genes related to DNA damage, stress responses, oxidative damage, apoptosis, and cell growth. RT-PCR results paralleled the Affymetrix results for four selected genes (HMOX1, DDIT4, GCLM, and CLK4). Our results show for the first time that VAN and CIN are antimutagenic against spontaneous mutations in mammalian (human) cells. These and other data lead us to propose that VAN and CIN may induce DNA damage that elicits recombinational DNA repair, which reduces spontaneous mutations.
authors.affiliation: The Nelson Institute of Environmental Medicine, New York University School of Medicine, 57 Old Forge Road, Tuxedo, NY 10987, USA.
authors.forename: Audrey A
authors.initials: AA
authors.lastname: King
authors.forename: Daniel T
authors.initials: DT
authors.lastname: Shaughnessy
authors.forename: Kanae
authors.initials: K
authors.lastname: Mure
authors.forename: Joanna
authors.initials: J
authors.lastname: Leszczynska
authors.forename: William O
authors.initials: WO
authors.lastname: Ward
authors.forename: David M
authors.initials: DM
authors.lastname: Umbach
authors.forename: Zongli
authors.initials: Z
authors.lastname: Xu
authors.forename: Danica
authors.initials: D
authors.lastname: Ducharme
authors.forename: Jack A
authors.initials: JA
authors.lastname: Taylor
authors.forename: David M
authors.initials: DM
authors.lastname: Demarini
authors.forename: Catherine B
authors.initials: CB
authors.lastname: Klein
mesh_terms: D000171:Acrolein / Q000031:analogs & derivatives* / Q000494:pharmacology; D048868:Adaptor Proteins, Signal Transducing; D016587:Antimutagenic Agents / Q000494:pharmacology*; D001547:Benzaldehydes / Q000494:pharmacology*; D002352:Carrier Proteins / Q000235:genetics; D002470:Cell Survival; D020552:Comet Assay; D004249:DNA Damage; D004260:DNA Repair; D004305:Dose-Response Relationship, Drug; D005421:Flavoring Agents / Q000494:pharmacology; D005786:Gene Expression Regulation; D045325:HCT116 Cells; D006801:Humans; D000070957:MutL Protein Homolog 1; D009154:Mutation; D009687:Nuclear Proteins / Q000235:genetics; D020411:Oligonucleotide Array Sequence Analysis; D015398:Signal Transduction
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D016587:Antimutagenic Agents; D001547:Benzaldehydes; D002352:Carrier Proteins; D005421:Flavoring Agents; C501486:MLH1 protein, human; D009687:Nuclear Proteins; D000171:Acrolein; C100058:vanillin; D000070957:MutL Protein Homolog 1; C012843:cinnamaldehyde