c-Met tyrosine kinase receptor expression and function in human and canine osteosarcoma cells.
To further characterize the role of hepatocyte growth factor-scatter factor (HGF-SF) and its receptor (c-Met) in osteosarcoma (OS), human OS cell lines with low (SAOS-2) and high (SAOS-LM2) metastatic potential, and cell lines derived from spontaneous canine OS were studied. All cell lines were evaluated for c-Met and HGF-SF expression and receptor activation using Northern, RT-PCR, and Western blot analyses, respectively. Functional activity of receptor-ligand interaction was measured using c-Met phosphorylation status, proliferation assays (anchorage-dependent and -independent), Matrigel invasion, modulation of urokinase plasminogen activator (uPA) expression, and cell dispersion (scattering). All cell lines exhibited steady-state mRNA expression of c-Met. The canine OS cell lines also expressed HGF-SF mRNA as determined by RT-PCR analysis. Western analysis showed c-Met protein expression and HGF-stimulated (human) or constitutive (canine) receptor autophosphorylation. Treatment with recombinant human HGF resulted in enhanced proliferation in 3 of 5 OS cell lines and enhanced colony formation in 2 of 5 OS cell lines. Matrigel invasion was significantly enhanced in 3 of the cell lines and uPA levels were significantly increased in the SAOS-2 cells following HGF treatment. Scattering was enhanced in both the SAOS-2 and SAOS-LM2 cells. These data support the involvement of c-Met and HGF-SF in the growth and progression of human and canine OS, and may offer new targets for the development of therapeutic strategies for OS.
MacEwen
E Gregory
EG
Department of Medical Sciences, School of Veterinary Medicine, University of Wisconsin-Madison, Madison, Wisconsin 53706, USA.
Kutzke
Jon
J
Carew
Jennifer
J
Pastor
Josep
J
Schmidt
Julie A
JA
Tsan
Rachel
R
Thamm
Douglas H
DH
Radinsky
Robert
R
D000818:Animals; D015152:Blotting, Northern; D015153:Blotting, Western; D002455:Cell Division; D003094:Collagen / Q000494:pharmacology; D004285:Dogs; D004338:Drug Combinations; D017228:Hepatocyte Growth Factor / Q000502:physiology; D006801:Humans; D007797:Laminin / Q000494:pharmacology; D008024:Ligands; D009362:Neoplasm Metastasis; D009693:Nucleic Acid Hybridization; D012516:Osteosarcoma / Q000378:metabolism*; D010766:Phosphorylation; D011509:Proteoglycans / Q000494:pharmacology; D019859:Proto-Oncogene Proteins c-met / Q000096:biosynthesis* / Q000502:physiology*; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014407:Tumor Cells, Cultured; D014568:Urokinase-Type Plasminogen Activator / Q000096:biosynthesis
D004338:Drug Combinations; D007797:Laminin; D008024:Ligands; D011509:Proteoglycans; D012333:RNA, Messenger; C059485:matrigel; D017228:Hepatocyte Growth Factor; D003094:Collagen; D019859:Proto-Oncogene Proteins c-met; D014568:Urokinase-Type Plasminogen Activator