c-Met tyrosine kinase receptor expression and function in human and canine osteosarcoma cells.

Abstract

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.

• Authors: E Gregory MacEwen, Jon Kutzke, Jennifer Carew, Josep Pastor, Julie A Schmidt, Rachel Tsan, Douglas H Thamm, Robert Radinsky
• Journal: Clinical & experimental metastasis (Clin Exp Metastasis) Vol. 20 Issue 5 Pg. 421-30 ( 2003) ISSN: 0262-0898 [Print] Netherlands
• PMID: 14524531 (Publication Type: Journal Article, Research Support, U.S. Gov't, P.H.S.)

Chemical References

• Drug Combinations
• Laminin
• Ligands
• Proteoglycans
• RNA, Messenger
• matrigel
• Hepatocyte Growth Factor
• Collagen
• Proto-Oncogene Proteins c-met
• Urokinase-Type Plasminogen Activator

Topics

• Animals
• Blotting, Northern
• Blotting, Western
• Cell Division
• Collagen (pharmacology)
• Dogs
• Drug Combinations
• Hepatocyte Growth Factor (physiology)
• Humans
• Laminin (pharmacology)
• Ligands
• Neoplasm Metastasis
• Nucleic Acid Hybridization
• Osteosarcoma (metabolism)
• Phosphorylation
• Proteoglycans (pharmacology)
• Proto-Oncogene Proteins c-met (biosynthesis, physiology)
• RNA, Messenger (metabolism)
• Reverse Transcriptase Polymerase Chain Reaction
• Tumor Cells, Cultured
• Urokinase-Type Plasminogen Activator (biosynthesis)