Novel Quantitation of Autocrine/Paracrine Stimulation of Cell Motility in Vitro and Metastasis
Abstract
Three established human breast cancer cell lines were analyzed and found to express the L1 adhesion/recognition molecule by immunofluorescence, RT-PCR, and western blotting. Protein expression levels correlated to their known metastatic abilities. Cells released a large soluble proteolytic L1 fragment and tiny L1-containing vesicles into culture media. Breast cancer cells express multiple integrin cell surface receptors that can bind to the released or shed L1. L1 proteolysis can be upregulated by a phorbol ester, indicating that this phenomenon can be modulated. A L1-shRNA retroviral vector was found that attenuated expression of breast cancer cells, and 3 retroviral vectors were constructed that cause expression of different length L1 ectodomain fragments. The chick embryo was found to be a sensitive system for studying breast cancer metastasis by using as few as 5,000 cells injected into the blood stream followed by recovery and culture of cells from the brain.
Document Details
- Document Type
- Technical Report
- Publication Date
- Sep 01, 2008
- Accession Number
- ADA510235
Entities
People
- Deni S. Galileo
Organizations
- University of Delaware