title: Lymphatic vessels assessment in feline mammary tumours.
abstract: BACKGROUND
  The lymphatic vessels play a crucial role in a variety of human cancers since tumour cell lymphatic invasion significantly influences prognosis. It is not known if pre-existing lymphatics are enough for tumour dissemination or de novo development is necessary. VEGFR-3 is an angiogenetic mediator for both lymphatic and blood vessels during embryonic development, and only for lymphatics after birth. VEGF is a mediator of both vasculogenesis and angiogenesis, regulates the growth of lymphatics in various experimental models, and is produced in many solid tumours. CD44 mediates hyaluronic acid (HA)-dependent cell adhesion: besides promoting invasion, this interaction also supports neoangiogenesis that indirectly stimulates tumour cell proliferation. The expression of VEGF-C (Vascular Endothelial Growth Factor-C), its receptor VEGFR-3 and CD44, were studied on feline mammary samples to assess the importance of lymphangiogenesis and lymphangiotrophism in neoplasia.
  
  
  METHODS
  Samples were taken from six normal mammary glands (NMG), ten benign (BT) and 32 malignant (MT) tumours. Immunohistochemical laminin/VEGFR-3 double stain, VEGF-C and CD44 stains were applied to 4 mum-thick sections, and their expression evaluated in intratumoral/extratumoral and intramammary/extramammary fields.
  
  
  RESULTS
  All groups revealed a higher number of lymphatics in the extratumoral/extramammary areas. VEGF-C expression in the epithelium paralleled the number of positive vessels in the NMG, BT and MT, whereas VEGF-C higher expression was noted in the intratumoral fields only in infiltrating MT. CD44 score was lower in extratumoral than intratumoral fields in tumours and showed a significant increase in extramammary/extratumoral fields from NMG to MT. Pearson test showed a significant and inversely proportional correlation between CD44 expression and the number of lymphatic vessels with VEGFR-3 in malignant infiltrating tumours.
  
  
  CONCLUSIONS
  The number of both VEGFR-3 positive and negative lymphatics in the extratumoral and extramammary stroma was significantly higher than intratumoral and intramammary fields respectively in the NMG, BT and MT. This suggests a scant biological importance of intratumoral lymphatics while their higher number is due to the concentration of existing vessels following compression of the extratumoral stroma in spite of a non demonstrable increase from NMG to MT. The tumour model employed provided no evidence of lymphangiogenesis, and metastasis in the regional lymph node develops following the spread through the pre-existing lymphatic network.
authors.affiliation: Department of Veterinary Public Health and Animal Pathology, Bologna, Italy. giuseppe.sarli@unibo.it
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Sarli
authors.forename: Francesco
authors.initials: F
authors.lastname: Sassi
authors.forename: Barbara
authors.initials: B
authors.lastname: Brunetti
authors.forename: Antonio
authors.initials: A
authors.lastname: Rizzo
authors.forename: Laura
authors.initials: L
authors.lastname: Diracca
authors.forename: Cinzia
authors.initials: C
authors.lastname: Benazzi
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002415:Cats; D018960:Hyaluronan Receptors / Q000378:metabolism; D042583:Lymphangiogenesis; D008207:Lymphatic Metastasis / Q000503:physiopathology*; D042601:Lymphatic Vessels / Q000378:metabolism*; D015674:Mammary Neoplasms, Animal / Q000378:metabolism*; D042582:Vascular Endothelial Growth Factor C / Q000378:metabolism; D040321:Vascular Endothelial Growth Factor Receptor-3 / Q000378:metabolism
chemical_list: D015415:Biomarkers; D018960:Hyaluronan Receptors; D042582:Vascular Endothelial Growth Factor C; D040321:Vascular Endothelial Growth Factor Receptor-3