Molecular Mechanisms in Compromised Endothelial Barrier during Breast Cancer Metastasis
Abstract
In this funding period (second of three years), we have completed the first specific as scheduled. We have successfully devised a novel assay system comprise of an engineered 3D vasculature network stably expressing a FRET-based biosensor for myosin light chain kinase (MLCK) activity. This set-up thus offers a very powerful assay system to directly study tumor invasion of the vascular system from the perspective of the endothelial cells. Using this assay, we have observed that endothelial cell MLCK signal is closely mediated by the interaction of the endothelial cells with the invading tumor. Our results implicated a biphasic endothelial signaling response to the interaction with metastatic tumor cells: (1) an initial, general, elevation of MLCK activity above baseline when the cancer cells crawl along the endothelial cell surface, (2) a more acute and marked increase in MLCK activity at the site of tumor entry during active invasion. We also observed that the majority of cancer cells enter the engineered vasculature system via the transcellular route (i.e. through individual endothelial cells), rather than through the paracellular mechanism (through cell-cell junctions), as conventionally believed. We are now actively characterizing the signal of MLCK as well as the reorganization of the actin-myosin network during this event.
Document Details
- Document Type
- Technical Report
- Publication Date
- Mar 01, 2008
- Accession Number
- ADA485716
Entities
People
- Teng-leong Chew
Organizations
- Northwestern University