title: Endocytosis of resveratrol via lipid rafts and activation of downstream signaling pathways in cancer cells.
abstract: trans-Resveratrol has been proposed to prevent tumor growth and to sensitize cancer cells to anticancer agents. Polyphenol entry into the cells has remained poorly understood. Here, we show that [(3)H]-resveratrol enters colon cancer cells (SW480, SW620, HT29) and leukemia U937 cells through a monensin (5-20 μmol/L) -sensitive process that suggests clathrin-independent endocytosis. Uptake of the molecule can be prevented by methyl-β-cyclodextrin (2-12 mg/mL), nystatin (12 ng/mL), and filipin (1 μg/mL), which all disrupt plasma membrane lipid rafts. Accordingly, radiolabeled resveratrol accumulates in sphingomyelin- and cholesterol-enriched cell fractions. Interestingly, extracellular signal-regulated kinases (ERK), c-Jun NH(2)-terminal kinases (JNK), and Akt also accumulate in lipid rafts on resveratrol exposure (IC(50) at 48 h ≈ 30 μmol/L in SW480 and U937 cells). In these rafts also, resveratrol promotes the recruitment, by the integrin α(V)β(3) (revealed by coimmunoprecipitation with an anti-integrin α(V)β(3) antibody), of signaling molecules that include the FAK (focal adhesion kinase), Fyn, Grb2, Ras, and SOS proteins. Resveratrol-induced activation of downstream signaling pathways and caspase-dependent apoptosis is prevented by endocytosis inhibitors, lipid raft-disrupting molecules, and the integrin antagonist peptide arginine-glycine-aspartate (500 nmol/L). Altogether, these data show the role played by lipid rafts in resveratrol endocytosis and activation of downstream pathways leading to cell death.
authors.affiliation: Inserm UMR866, 6 Boulevard Gabriel, 21000 Dijon, France.
authors.forename: Didier
authors.initials: D
authors.lastname: Colin
authors.forename: Emeric
authors.initials: E
authors.lastname: Limagne
authors.forename: Sylvie
authors.initials: S
authors.lastname: Jeanningros
authors.forename: Arnaud
authors.initials: A
authors.lastname: Jacquel
authors.forename: Gérard
authors.initials: G
authors.lastname: Lizard
authors.forename: Anne
authors.initials: A
authors.lastname: Athias
authors.forename: Philippe
authors.initials: P
authors.lastname: Gambert
authors.forename: Aziz
authors.initials: A
authors.lastname: Hichami
authors.forename: Norbert
authors.initials: N
authors.lastname: Latruffe
authors.forename: Eric
authors.initials: E
authors.lastname: Solary
authors.forename: Dominique
authors.initials: D
authors.lastname: Delmas
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D015153:Blotting, Western; D003110:Colonic Neoplasms / Q000188:drug therapy*; D004705:Endocytosis / Q000502:physiology*; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D005372:Filipin / Q000494:pharmacology; D005434:Flow Cytometry; D051417:Focal Adhesion Kinase 1 / Q000378:metabolism; D051380:GRB2 Adaptor Protein / Q000378:metabolism; D006801:Humans; D007124:Immunoenzyme Techniques; D047468:Immunoprecipitation; D039302:Integrin alphaVbeta3 / Q000276:immunology / Q000378:metabolism; D021962:Membrane Microdomains / Q000502:physiology*; D009761:Nystatin / Q000494:pharmacology; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D051076:Proto-Oncogene Proteins c-fyn / Q000378:metabolism; D000077185:Resveratrol; D015398:Signal Transduction / Q000187:drug effects*; D020664:Son of Sevenless Proteins / Q000378:metabolism; D013267:Stilbenes / Q000494:pharmacology*; D014407:Tumor Cells, Cultured; D047392:beta-Cyclodextrins / Q000494:pharmacology; D018631:ras Proteins / Q000378:metabolism
chemical_list: D000900:Anti-Bacterial Agents; D000911:Antibodies, Monoclonal; D000972:Antineoplastic Agents, Phytogenic; D051380:GRB2 Adaptor Protein; C495948:GRB2 protein, human; D039302:Integrin alphaVbeta3; D020664:Son of Sevenless Proteins; D013267:Stilbenes; D047392:beta-Cyclodextrins; C108732:methyl-beta-cyclodextrin; D009761:Nystatin; D005372:Filipin; D051417:Focal Adhesion Kinase 1; C496050:PTK2 protein, human; D051076:Proto-Oncogene Proteins c-fyn; D051057:Proto-Oncogene Proteins c-akt; D048049:Extracellular Signal-Regulated MAP Kinases; D018631:ras Proteins; D000077185:Resveratrol