Related Experiment Videos
Requirement of regulated activation of Ras for response of MDCK cells to hepatocyte growth factor/scatter factor
1Department of Life Science, Faculty of Bioscience and Biotechnology, Tokyo Institute of Technology, Nagatsuta, Midori-ku, Yokohama, 226-8501, Japan.
Abstract:
Hepatocyte growth factor/scatter factor (HGF/SF) induces cell scattering, migration, and branching tubule formation of MDCK cells. To examine the role of the Ras protein in the HGF/SF-induced responses, we constructed MDCK cell clones expressing either inducible dominant-negative Ras or constitutively activated Ras and analyzed their effects on responses of cells to HGF/SF. Induced expression of dominant-negative Ras prevented cell dissociation required for cell scattering, migration, and cystic formation as well as branching morphology required for branching tubule formation. Constitutively activated Ras induced cell dissociation, but not a scattered fibroblastic morphology even in the presence of HGF/SF. MDCK cells expressing constitutively activated Ras migrated at a level similar to that of wild-type MDCK cells stimulated by HGF/SF. MDCK cells expressing constitutively activated Ras showed disorganized growth in three-dimensional culture and did not form the branching tubule structures. These results indicate that activation of the Ras protein is essential for the cell scattering, migration, and branching tubule formation of MDCK cells induced by HGF/SF, and a properly regulated activation is required for some stages of the HGF/SF-induced responses of MDCK cells.
Insights
Ras protein activation is essential for hepatocyte growth factor/scatter factor (HGF/SF) to induce cell scattering, migration, and tubule formation in MDCK cells. Proper Ras regulation is crucial for these HGF/SF-driven cellular responses.
Area of Science:
- Cell Biology
- Molecular Biology
- Biochemistry
Background:
- Hepatocyte growth factor/scatter factor (HGF/SF) is known to induce cell scattering, migration, and tubule formation in Madin-Darby Canine Kidney (MDCK) cells.
- The precise role of the Ras signaling pathway in mediating these HGF/SF-induced cellular behaviors remains to be fully elucidated.
Purpose of the Study:
- To investigate the role of Ras protein activation in HGF/SF-induced cell scattering, migration, and branching tubule formation in MDCK cells.
- To determine if dominant-negative or constitutively activated Ras impacts HGF/SF-mediated cellular responses.
Main Methods:
- Generation of MDCK cell clones with inducible dominant-negative Ras or constitutively activated Ras.
- Analysis of HGF/SF-induced cell scattering, migration, and tubule formation in these engineered cell lines.
- Comparison of cellular responses between engineered cells and wild-type MDCK cells.
Main Results:
- Inducible dominant-negative Ras expression blocked HGF/SF-induced cell dissociation, scattering, migration, and branching tubule formation.
- Constitutively activated Ras induced cell dissociation but not scattered morphology, and migration levels were similar to HGF/SF-stimulated wild-type cells.
- Cells with constitutively activated Ras exhibited disorganized growth and failed to form branching tubules in 3D culture.
Conclusions:
- Ras protein activation is essential for HGF/SF-induced cell scattering, migration, and branching tubule formation in MDCK cells.
- Properly regulated Ras activation is required for specific stages of HGF/SF-mediated cellular responses, highlighting the importance of controlled signaling.