title: Leukocyte recruitment in colon cancer: role of cell adhesion molecules, nitric oxide, and transforming growth factor beta1.
abstract: OBJECTIVE
  A deficient leukocyte recruitment has been suggested in tumor vasculature, but little is known about the underlying molecular mechanism. To characterize leukocyte-endothelium interaction in experimental colon cancer, quantify the main endothelial cell adhesion molecules (CAMs), and evaluate the effect of tumor-derived products.
  
  
  METHODS
  Leukocyte recruitment was assessed by intravital videomicroscopy in mice bearing HT29-derived tumors. Endothelial CAMs were measured using the dual-radiolabeled antibody technique. The role of molecules mediating leukocyte rolling (P-, E-, and L-selectin) or adhesion (intercellular adhesion molecule 1 [ICAM-1] and vascular cell adhesion molecule 1 [VCAM-1]) carcinoembryonic antigen (CEA), and transforming growth factor (TGF) beta1 was assessed through immunoblockade, whereas participation of nitric oxide (NO) and cyclooxygenase (COX) metabolites were evaluated by means of nonselective and selective inhibition.
  
  
  RESULTS
  Basal and lipopolysaccharide-stimulated leukocyte rolling and adhesion were markedly reduced in tumor vasculature. ICAM-1 immunoblockade prevented leukocyte adhesion in both tumor and nontumor microvessels. Neither baseline nor LPS-induced endothelial ICAM-1, P-, and E-selectin expression in tumors were reduced with respect to nontumor vasculature. Although VCAM-1 expression was reduced in tumor endothelium, immunoneutralization of VCAM-1 failed to reverse LPS-induced leukocyte recruitment in this setting. CEA immunoblockade and COX inhibition did not modify the deficient leukocyte rolling. Nonselective NO inhibition partially reversed the defective adhesion response in tumor microvessels. Finally, TGF-beta1 immunoblockade partially and selectively restored impaired leukocyte rolling and adhesion in tumor microvessels.
  
  
  CONCLUSIONS
  Impaired leukocyte recruitment in tumor vasculature cannot be attributed to a depressed expression of the main CAMs. Selective restoration after NO inhibition and TGF-beta1 immunoblockade suggests involvement of both molecules in this phenomenon.
authors.affiliation: Gastroenterology Department, Institut de Malalties Digestives, Hospital Clínic, Institut d'Investigacions Biomèdiques Agust Pi i Sunyer, University of Barcelona, Barcelona, Spain.
authors.forename: Xavier
authors.initials: X
authors.lastname: Bessa
authors.forename: J Ignasi
authors.initials: JI
authors.lastname: Elizalde
authors.forename: Francesc
authors.initials: F
authors.lastname: Mitjans
authors.forename: Virgínia
authors.initials: V
authors.lastname: Piñol
authors.forename: Rosa
authors.initials: R
authors.lastname: Miquel
authors.forename: Julián
authors.initials: J
authors.lastname: Panés
authors.forename: Jaime
authors.initials: J
authors.lastname: Piulats
authors.forename: Josep M
authors.initials: JM
authors.lastname: Piqué
authors.forename: Antoni
authors.initials: A
authors.lastname: Castells
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D002272:Carcinoembryonic Antigen / Q000378:metabolism; D002448:Cell Adhesion / Q000276:immunology; D002450:Cell Communication / Q000276:immunology; D002634:Chemotaxis, Leukocyte / Q000502:physiology; D003110:Colonic Neoplasms / Q000098:blood supply / Q000276:immunology* / Q000378:metabolism; D019040:E-Selectin / Q000276:immunology / Q000378:metabolism; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism; D019073:HT29 Cells; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000276:immunology / Q000378:metabolism*; D019041:L-Selectin / Q000276:immunology / Q000378:metabolism; D007962:Leukocytes / Q000166:cytology* / Q000378:metabolism; D051379:Mice; D008819:Mice, Nude; D009569:Nitric Oxide / Q000378:metabolism*; D019007:P-Selectin / Q000276:immunology / Q000378:metabolism; D011451:Prostaglandin-Endoperoxide Synthases / Q000378:metabolism; D016212:Transforming Growth Factor beta / Q000378:metabolism*; D053773:Transforming Growth Factor beta1; D019010:Vascular Cell Adhesion Molecule-1 / Q000276:immunology / Q000378:metabolism*
chemical_list: D000911:Antibodies, Monoclonal; D002272:Carcinoembryonic Antigen; D019040:E-Selectin; D019007:P-Selectin; C508960:TGFB1 protein, human; C508961:Tgfb1 protein, mouse; D016212:Transforming Growth Factor beta; D053773:Transforming Growth Factor beta1; D019010:Vascular Cell Adhesion Molecule-1; D018799:Intercellular Adhesion Molecule-1; D019041:L-Selectin; D009569:Nitric Oxide; D011451:Prostaglandin-Endoperoxide Synthases