title: Auraptene, a citrus coumarin, inhibits 12-O-tetradecanoylphorbol-13-acetate-induced tumor promotion in ICR mouse skin, possibly through suppression of superoxide generation in leukocytes.
abstract: Coumarin-related compounds, auraptene and umbelliferone, have been isolated from the cold-pressed oil of natsumikan (Citrus natsudaidai HAYATA), and tested as inhibitors of tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced Epstein-Barr virus activation in Raji cells. The 50% inhibitory concentration (IC50) of auraptene (18 microM) was almost equal to that of genistein. Umbelliferone, which lacks a geranyloxyl group present in auraptene, was less active (IC50 = 450 microM). In a two-stage carcinogenesis experiment with 7,12-dimethylbenz[a] anthracene (topical application at 0.19 mumol) and TPA (topical application at 1.6 nmol) in ICR mouse skin, topical application of auraptene (at 160 nmol) significantly reduced tumor incidence and the numbers of tumors per mouse by 27% (P < 0.01) and 23% (P < 0.05), respectively. Auraptene at a concentration of 50 microM markedly suppressed superoxide (O2-) generation induced by 100 microM TPA in differentiated human promyelocytic HL-60 cells. Having no O2(-)-scavenging potential, auraptene may inhibit the multicomponent NADPH oxidase system. Inhibition of intracellular hydroperoxide formation in differentiated HL-60 cells by auraptene was also confirmed by flow-cytometric analysis using 2',7'-dichlorofluorescein diacetate as a fluorescence probe. Quantitative analyses using high-performance liquid chromatography showed the occurrence of auraptene not only in both the peels and sarcocarps of natsumikan, but also in those of hassaku orange (C. hassaku) and grapefruit (C. paradisi), and even in their bottled fresh juice form. These results indicate that auraptene is a chemopreventer of skin tumorigenesis, and implies that suppression of leukocyte activation might be the mechanism through which it inhibits tumor promotion.
authors.affiliation: Department of Biotechnological Science, Faculty of Biology-Oriented Science and Technology, Kinki University, Wakayama.
authors.forename: A
authors.initials: A
authors.lastname: Murakami
authors.forename: W
authors.initials: W
authors.lastname: Kuki
authors.forename: Y
authors.initials: Y
authors.lastname: Takahashi
authors.forename: H
authors.initials: H
authors.lastname: Yonei
authors.forename: Y
authors.initials: Y
authors.lastname: Nakamura
authors.forename: Y
authors.initials: Y
authors.lastname: Ohto
authors.forename: H
authors.initials: H
authors.lastname: Ohigashi
authors.forename: K
authors.initials: K
authors.lastname: Koshimizu
mesh_terms: D015127:9,10-Dimethyl-1,2-benzanthracene; D000818:Animals; D016588:Anticarcinogenic Agents / Q000627:therapeutic use*; D002273:Carcinogens; D002460:Cell Line; D002957:Citrus / Q000737:chemistry*; D003374:Coumarins / Q000032:analysis / Q000627:therapeutic use*; D005260:Female; D005434:Flow Cytometry; D005456:Fluorescent Dyes; D006801:Humans; D015473:Leukemia, Promyelocytic, Acute / Q000378:metabolism; D007962:Leukocytes / Q000187:drug effects / Q000378:metabolism*; D051379:Mice; D008813:Mice, Inbred ICR; D012878:Skin Neoplasms / Q000139:chemically induced / Q000517:prevention & control*; D013481:Superoxides / Q000378:metabolism*; D013755:Tetradecanoylphorbol Acetate; D014468:Umbelliferones / Q000627:therapeutic use
chemical_list: D016588:Anticarcinogenic Agents; D002273:Carcinogens; D003374:Coumarins; D005456:Fluorescent Dyes; D014468:Umbelliferones; D013481:Superoxides; D015127:9,10-Dimethyl-1,2-benzanthracene; C031477:7-hydroxycoumarin; C105832:aurapten; D013755:Tetradecanoylphorbol Acetate