Spherical Invasion Assay: A Novel Method to Measure Invasion of Cancer Cells.

Area of Science:

• Cancer Biology
• Cellular Invasion Assays
• Drug Discovery

Background:

• Tumor cell invasion is critical for metastasis.
• Traditional Boyden chamber assays have limitations in physiological relevance and gradient control.
• There is a need for alternative methods to validate cancer cell invasion studies.

Purpose of the Study:

• To develop and validate a Spherical Invasion Assay (SIA) for measuring cancer cell invasion.
• To assess the anti-invasive activity of the Src kinase inhibitor PP2 in A549 non-small cell lung carcinoma cells using the SIA.
• To compare SIA results with traditional Boyden chamber assays.

Main Methods:

• A two-step SIA protocol involving seeding cancer cells in Matrigel, followed by a second Matrigel overlay.
• Treatment of A549 cells with the Src kinase inhibitor PP2.
• Quantification of cell invasion using phase-contrast microscopy and ImageJ software.

Main Results:

• The SIA protocol successfully measured the anti-invasive activity of PP2 in A549 cells.
• Results from the SIA correlated well with those from Boyden chamber assays.
• The SIA demonstrated potential for drug screening and studying invasion pathways.

Conclusions:

• The SIA is a viable and physiologically relevant method for assessing cancer cell invasion.
• The SIA can be used to validate findings from traditional invasion assays.
• The SIA has broad applications in cancer research, including drug development and pathway elucidation.