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Isolating Hair Follicle Stem Cells and Epidermal Keratinocytes from Dorsal Mouse Skin
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HGF-induced DU145 cell scatter assay.

Sally T Fram1, Claire M Wells, Gareth E Jones

  • 1Randall Division of Cell and Molecular Biophysics, King’s College London, London, UK.

Methods in Molecular Biology (Clifton, N.J.)
|July 13, 2011
PubMed
Summary

We present an optimized protocol for studying epithelial mesenchymal transition (EMT) using the DU145 human prostate cancer cell line. This method effectively monitors changes in cell adhesion, migration, and actin cytoskeleton organization during HGF-induced cell scattering.

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Area of Science:

  • Cell Biology
  • Cancer Research
  • Biochemistry

Background:

  • Epithelial mesenchymal transition (EMT) is crucial for development and cancer invasion.
  • EMT involves loss of cell adhesion and increased cell migration, dependent on actin cytoskeleton reorganization.
  • Hepatocyte growth factor (HGF) is known to induce EMT-like cell scattering in certain cell types.

Purpose of the Study:

  • To describe an optimized protocol for an HGF-induced DU145 cell scattering assay.
  • To establish a human cell model for studying EMT.
  • To facilitate monitoring of cytoskeletal dynamics, cell adhesion, and migration.

Main Methods:

  • Utilized the human prostate cancer cell line DU145.
  • Applied Hepatocyte Growth Factor (HGF) to induce cell scattering.
  • Optimized protocols for assay conduct and analysis.
  • Monitored actin cytoskeleton organization, cell-cell adhesions, and cell migration.

Main Results:

  • Developed a robust model of HGF-induced cell scattering in DU145 cells.
  • Demonstrated the utility of this model for observing EMT-related cellular changes.
  • Successfully monitored dynamic alterations in actin cytoskeleton, cell junctions, and migratory behavior.

Conclusions:

  • The HGF-induced DU145 cell scattering assay provides a valuable model for studying EMT in human cells.
  • This assay is effective for analyzing changes in cell migration and cytoskeletal dynamics.
  • The protocol enables detailed investigation of molecular events underlying EMT in cancer research.