title: The potential lymphangiogenic effects of hepatocyte growth factor/scatter factor in vitro and in vivo.
abstract: Lymphangiogenesis is key to the lymphatic spread of cancer cells. The current study examined the potential effect of hepatocyte growth factor (HGF), a factor known to have strong biological effects on endothelial cells, on the lymphangiogenic function of endothelial cells and the formation of lymphatic vessels using both in vitro and in vivo models. Human endothelial cells that have lymphatic characteristics, human prostate and breast cancer cells PC-3 and MDA MB 231, were used. Expression of lymphatic markers, podoplanin, Prox-1, vascular endothelial growth factor receptor 3 (VEGF-R3) and LYVE-1 was determined using reverse transcription polymerase reaction and quantitative PCR. In nude mice prostate and breast xenograft tumour models, either HGF or an HGF-producing fibroblast cell line MRC-5 was given with or without the HGF antagonist, NK4. The lymphangiogenic marker and lymphatic vessels in tumour tissues were also assessed using quantitative PCR and immunohistochemistry, respectively. In the mice tumour models, infusion of rhHGF significantly increased the levels of podoplanin and LYVE-1 in the tumour (p=0.05 for podoplanin and p<0.05 for LYVE-1 vs. without HGF in the prostate tumour model, p<0.05 for podoplanin and p<0.01 for LYVE-1 vs. without HGF for the breast tumour model; p<0.05 for podoplanin and p<0.01 for LYVE-1 vs. without HGF in the breast tumour model). The increased level of LYVE-1 transcript was supported by an increase in the number of LYVE-1-positive lymphatic vessels in tumours, using immunohistochemical analysis. Co-injection of MRC5 cells also increased the levels of LYVE-1 and number of LYVE-1-positive vessels in tumour tissues. The effects of HGF and MRC5 were significantly reduced by the HGF antagonist, NK4. In the in vitro models, rhHGF significantly increased the level of both podoplanin and LYVE-1, as shown by quantitative PCR analysis. Hepatocyte growth factor has potential lymphangiogenic activities, and this may have important implications in the nodal spread of cancer cells.
authors.affiliation: Metastasis and Angiogenesis Research Group, Wales College of Medicine, Cardiff University, Heath Park, Cardiff CF4 4XN, UK. jiangw@cf.ac.uk
authors.forename: Wen G
authors.initials: WG
authors.lastname: Jiang
authors.forename: Gaynor
authors.initials: G
authors.lastname: Davies
authors.forename: Tracey A
authors.initials: TA
authors.lastname: Martin
authors.forename: Christian
authors.initials: C
authors.lastname: Parr
authors.forename: Gareth
authors.initials: G
authors.lastname: Watkins
authors.forename: Robert E
authors.initials: RE
authors.lastname: Mansel
authors.forename: Malcolm D
authors.initials: MD
authors.lastname: Mason
mesh_terms: D000818:Animals; D002460:Cell Line; D045744:Cell Line, Tumor; D004305:Dose-Response Relationship, Drug; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D005260:Female; D015870:Gene Expression / Q000187:drug effects; D006023:Glycoproteins / Q000235:genetics / Q000378:metabolism; D017228:Hepatocyte Growth Factor / Q000494:pharmacology*; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D042583:Lymphangiogenesis / Q000187:drug effects*; D042601:Lymphatic Vessels / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism; D026901:Membrane Transport Proteins; D051379:Mice; D008819:Mice, Nude; D008934:Mitogens / Q000494:pharmacology; D009368:Neoplasm Transplantation; D009374:Neoplasms, Experimental / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014183:Transplantation, Heterologous; D025521:Tumor Suppressor Proteins; D040321:Vascular Endothelial Growth Factor Receptor-3 / Q000235:genetics / Q000378:metabolism; D033921:Vesicular Transport Proteins; D000098551:Prospero-Related Homeobox 1 Protein; D000098853:Podoplanin
chemical_list: D006023:Glycoproteins; D017228:Hepatocyte Growth Factor; D018398:Homeodomain Proteins; D008562:Membrane Glycoproteins; D026901:Membrane Transport Proteins; D008934:Mitogens; D025521:Tumor Suppressor Proteins; D040321:Vascular Endothelial Growth Factor Receptor-3; D033921:Vesicular Transport Proteins; D000098551:Prospero-Related Homeobox 1 Protein; C490441:Pdpn protein, mouse; D000098853:Podoplanin; C119248:LYVE1 protein, human; C117468:PDPN protein, human; C492273:LYVE1 protein, mouse