PAI-1 Gene as a Target for Breast Cancer Therapy
Abstract
The purpose of work in year 03 of this study was to continue to define the in vitro growth characteristics of stable cell lines of human breast carcinoma cells (developed in year 01 and continuing into year 02) that synthesized varying levels of plasminogen activator inhibitor type-1 (Pai-1) as a result of transfection with expression vectors bearing PAI-1 cDNA inserts cloned in the sense and antisense orientations. A complete panel of 32 such genetically-engineered epithelial cell lines was created and each of these were assessed with regard to their ability to (1) locomote across a planar surface following scrape-injury of confluent monolayers (directed cell migration) and (2) exhibit invasive growth behavior in three-dimensional motility chamber assays. PAI-1 expression was required for epithelial cell migration in directed assays since antisense targeting effectively suppressed induced cell locomotion. A unique expression vector consisting of a PAI-1-GFP chimeric protein driven by PAI-1 promoter sequences was used to conclusively demonstrate PAI-1 deposition into cellular migration tracks. A "window" of PAI-1 expression was found to be necessary to support optimal breast cancer locomotion and invasive migration. Work in the final year will test this hypothesis in vivo.
Document Details
- Document Type
- Technical Report
- Publication Date
- Apr 01, 2001
- Accession Number
- ADA392548
Entities
People
- Paul J. Higgins
Organizations
- Albany Medical College