C-Met tyrosine kinase receptor expression and function in human and canine osteosarcoma cells
Copyright policy )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.
- The University of Texas System United States
- University of Wisconsin–Madison United States
- University of Wisconsin–Oshkosh United States
- The University of Texas MD Anderson Cancer Center United States
- University of Wisconsin System United States
Osteosarcoma, Hepatocyte Growth Factor, Reverse Transcriptase Polymerase Chain Reaction, Blotting, Western, Nucleic Acid Hybridization, Proto-Oncogene Proteins c-met, Blotting, Northern, Ligands, Drug Combinations, Dogs, Tumor Cells, Cultured, Animals, Humans, Proteoglycans, Collagen, Laminin, RNA, Messenger, Neoplasm Metastasis, Phosphorylation, Cell Division
Osteosarcoma, Hepatocyte Growth Factor, Reverse Transcriptase Polymerase Chain Reaction, Blotting, Western, Nucleic Acid Hybridization, Proto-Oncogene Proteins c-met, Blotting, Northern, Ligands, Drug Combinations, Dogs, Tumor Cells, Cultured, Animals, Humans, Proteoglycans, Collagen, Laminin, RNA, Messenger, Neoplasm Metastasis, Phosphorylation, Cell Division
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