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In vitro Cell Migration and Invasion Assays
Published on: June 1, 2014
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Model Systems to Investigate NOX-Dependent Cell Migration and Invasiveness
Howard E Boudreau1, Thomas L Leto2
1Laboratory of Clinical Immunology and Microbiology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, USA.
Methods in Molecular Biology (Clifton, N.J.)
|June 8, 2019
Summary
Reactive oxygen species (ROS), generated by NADPH oxidase (NOX) enzymes, are key drivers of cell migration and invasion. This study details methods to investigate NOX enzyme roles in cell motility, aiding cancer metastasis research.
Area of Science:
- Cell Biology
- Biochemistry
- Cancer Research
Background:
- Reactive oxygen species (ROS) are increasingly recognized for their critical roles in cellular processes, including migration and invasion.
- The NADPH oxidase (NOX) enzyme family is a significant source of ROS in various cell types during migration.
- NOX enzymes are implicated in wound repair, epithelial barrier maintenance, and notably, in promoting metastatic cell migration and invasiveness.
Purpose of the Study:
- To outline and detail three distinct in vitro assays for investigating the involvement of NOX enzymes in cell motility.
- To provide experimental approaches for delineating the specific roles of NOX enzymes in cell migration and invasion.
- To facilitate research into the mechanisms of cancer metastasis by understanding NOX-mediated cell migration.
Main Methods:
- Scratch-wound repair assay to assess cell migration in response to a simulated wound.
- Matrigel invasion assay to evaluate the ability of cells to invade through an extracellular matrix barrier.
- Cell culture insert assay to measure migration from defined monolayer boundaries.
Main Results:
- The presented assays allow for the examination of NOX enzyme involvement in cell motility under various experimental conditions.
- These methods enable the manipulation of endogenous or overexpressed NOX enzymes to elucidate their specific functions.
- The assays provide a comprehensive toolkit for dissecting the contribution of NOX-derived ROS to cell migration and invasion.
Conclusions:
- The described in vitro assays are valuable tools for studying the complex roles of NOX enzymes in cell migration.
- Understanding NOX enzyme function in cell motility is crucial for developing therapeutic strategies against diseases involving aberrant cell migration, such as cancer metastasis.
- These methodologies support further research into the biochemical pathways linking NOX activity to cell invasion.
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