title: Development of a quantitative bioassay to assess preventive compounds against inflammation-based carcinogenesis.
abstract: Reducing cancer incidence and mortality by use of cancer-chemopreventive agents is an important goal. We have established an in vitro bioassay that is able to screen large numbers of candidate chemicals that are positive for prevention of inflammation-related carcinogenesis. To accomplish this we have added candidate chemicals or vehicles and freshly isolated, fluorescent dye-labeled inflammatory cells that were overlaid on TNF-alpha-stimulated mouse endothelial cells in a 96-well plate. Inhibition of inflammatory cell attachment to the endothelial cells by the chemicals was quantified by the intensity of fluorescence from the adherent inflammatory cells after removing unattached cells. Using this assay, we selected two chemicals, auraptene and turmerones, for further study. As an in vivo test, diets containing these test chemicals were administered to mice with a piece of foreign body, gelatin sponge, that had been implanted to cause inflammation, and we found that the number of inflammatory cells that infiltrated into the subcutaneously implanted gelatin sponge was reduced compared to that found in the mice fed with a control diet. Moreover, diets containing either of the two chemicals prevented inflammation-based carcinogenesis in a mouse model. We found that the compounds reduced not only the number of infiltrating cells but also the expression of inducible nitric oxide synthase (iNOS) or formation of 8-hydroxy-2'-deoxyguanine (8-OHdG) in the infiltrated cells. Moreover, both compounds but not controls sustained the reducing activity in the inflammatory lesion, and this finding was confirmed by using non-invasive in vivo electron spin resonance. The newly established in vitro screening assay will be useful for finding biologically effective chemopreventive agents against inflammation-related carcinogenesis.
authors.affiliation: Division of Cancer Development System, National Cancer Center Research Institute, Tokyo 104-0045, Japan.
authors.forename: Kunishige
authors.initials: K
authors.lastname: Onuma
authors.forename: Yuka
authors.initials: Y
authors.lastname: Suenaga
authors.forename: Ryuta
authors.initials: R
authors.lastname: Sakaki
authors.forename: Satoshi
authors.initials: S
authors.lastname: Yoshitome
authors.forename: Yu
authors.initials: Y
authors.lastname: Sato
authors.forename: Satomi
authors.initials: S
authors.lastname: Ogawara
authors.forename: Saori
authors.initials: S
authors.lastname: Suzuki
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Kuramitsu
authors.forename: Hidekatsu
authors.initials: H
authors.lastname: Yokoyama
authors.forename: Akira
authors.initials: A
authors.lastname: Murakami
authors.forename: Junichi
authors.initials: J
authors.lastname: Hamada
authors.forename: Garth L
authors.initials: GL
authors.lastname: Nicolson
authors.forename: Masanobu
authors.initials: M
authors.lastname: Kobayashi
authors.forename: Junichi
authors.initials: J
authors.lastname: Fujii
authors.forename: Futoshi
authors.initials: F
authors.lastname: Okada
mesh_terms: D000818:Animals; D016588:Anticarcinogenic Agents / Q000627:therapeutic use; D001681:Biological Assay / Q000379:methods*; D002448:Cell Adhesion; D003374:Coumarins / Q000008:administration & dosage / Q000627:therapeutic use; D042783:Endothelial Cells / Q000187:drug effects* / Q000276:immunology; D005260:Female; D005354:Fibrosarcoma / Q000139:chemically induced / Q000188:drug therapy; D005453:Fluorescence; D007150:Immunohistochemistry / Q000379:methods*; D007249:Inflammation / Q000517:prevention & control*; D007659:Ketones / Q000008:administration & dosage / Q000627:therapeutic use; D008748:Methylcholanthrene / Q000633:toxicity; D051379:Mice; D008810:Mice, Inbred C57BL; D009374:Neoplasms, Experimental; D010938:Plant Oils / Q000627:therapeutic use; D012717:Sesquiterpenes; D014050:Toluene / Q000008:administration & dosage / Q000031:analogs & derivatives / Q000627:therapeutic use; D014407:Tumor Cells, Cultured; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
chemical_list: D016588:Anticarcinogenic Agents; D003374:Coumarins; D007659:Ketones; D010938:Plant Oils; D012717:Sesquiterpenes; D014409:Tumor Necrosis Factor-alpha; C078098:ar-turmerone; D014050:Toluene; D008748:Methylcholanthrene; C105832:aurapten