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Updated: Feb 22, 2026
Characterization of Cell Membrane Extensions and Studying Their Roles in Cancer Cell Adhesion Dynamics
Published on: March 26, 2018
A Screening System for Evaluating Cell Extension Formation, Collagen Compaction, and Degradation in Drug Discovery
Asuka Yuda1, Christopher A McCulloch1
11 Matrix Dynamics Group, Faculty of Dentistry, University of Toronto, Toronto, ON, Canada.
Abstract:
The generation of cell extensions is critical for matrix remodeling in tissue invasion by cancer cells, but current methods for identifying molecules that regulate cell extension formation and matrix remodeling are not well adapted for screening purposes. We applied a grid-supported, floating collagen gel system (~100 Pa stiffness) to examine cell extension formation, collagen compaction, and collagen degradation in a single assay. With the use of cultured diploid fibroblasts, a fibroblast cell line, and two cancer cell lines, we found that compared with attached collagen gels (~2800 Pa), the mean number and length of cell extensions were respectively greater in the floating gels. In assessing specific processes in cell extension formation, compared with controls, the number of cell extensions was reduced by latrunculin B, β1 integrin blockade, and a formin FH2 domain inhibitor. Screening of a kinase inhibitor library (480 compounds) with the floating gel assay showed that compared with vehicle-treated cells, there were large reductions of collagen compaction, pericellular collagen degradation, and number of cell extensions after treatment with SB431542, SIS3, Fasudil, GSK650394, and PKC-412. These data indicate that the grid-supported floating collagen gel model can be used to screen for inhibitors of cell extension formation and critical matrix remodeling events associated with cancer cell invasion.
Insights
A new floating collagen gel assay effectively screens for molecules that inhibit cancer cell extension formation and matrix remodeling, crucial for tissue invasion. This method aids in discovering potential anti-cancer therapeutics.
Area of Science:
- Biophysics
- Cell Biology
- Cancer Research
Background:
- Cell extension formation and extracellular matrix remodeling are vital for cancer cell invasion.
- Existing screening methods for regulators of these processes are limited.
- A novel assay is needed to efficiently identify molecules impacting cell invasion.
Purpose of the Study:
- To develop and validate a grid-supported, floating collagen gel assay for screening cell extension formation and matrix remodeling.
- To identify inhibitors of cell extension formation and matrix remodeling using the developed assay.
Main Methods:
- A grid-supported, floating collagen gel system (~100 Pa stiffness) was used to assess cell extension formation, collagen compaction, and degradation.
- Cultured diploid fibroblasts, a fibroblast cell line, and two cancer cell lines were utilized.
- A kinase inhibitor library (480 compounds) was screened using the floating gel assay.
Main Results:
- Floating collagen gels supported greater cell extension formation compared to attached gels (~2800 Pa).
- Latrunculin B, β1 integrin blockade, and a formin FH2 domain inhibitor reduced cell extension numbers.
- SB431542, SIS3, Fasudil, GSK650394, and PKC-412 significantly reduced collagen compaction, degradation, and cell extension numbers.
Conclusions:
- The grid-supported floating collagen gel model is a robust platform for screening inhibitors of cell extension formation.
- This assay effectively identifies modulators of matrix remodeling events critical for cancer cell invasion.
- The developed model facilitates the discovery of potential therapeutic agents targeting cancer metastasis.
