Characterization of Breast Cancer Cell Death Induced by Interferons and Retinoids.
Abstract
We have previously observed that a combination of human Interferon-beta (IFN) and all-trans retinoic acid (RA) induces superior growth inhibitory responses in several human tumor cells. In particular this combination induces cell death similar to apoptosis in vitro, which could not be observed with individual agents. Preliminary studies identified no changes in the levels of known regulators of cell death such as p53, cyclin D and Bc12. Thus it is important to understand the cell growth suppressive mechanisms and the genes that mediate these effects. We hypothesized that the observed cell death might be mediated by either novel or hither to unimplicated gene products. Therefore, the major aim of this project is to identify the gene products that mediate the growth inhibitory/ cell death inducing activities of the combination of IFN and RA in human tumor cells. To directly identify these gene products we have proposed to employ a genetic technique that utilizes anti-sense knock-out as the strategy. In this technique, cells are transfected with cDNA libraries cloned in antisense orientation in an episomal vector and then are challenged to survive the effects induced by human IFN-beta and RA. The cDNA libraries were prepared from BT-20 human breast carcinoma cell line that was treated with the IFN/RA combination for various lengths of time. The surviving cells will carry an antisense product that inhibits the function of an endogenous gene thereby blocking its growth inhibitory functions. By preparing the Hirt DNAs, the episome can be isolated from the cells and can be used to transform bacterial Sequencing of the episome identified in bacterial screening would permit the identification of the gene products.
Document Details
- Document Type
- Technical Report
- Publication Date
- Jul 01, 1997
- Accession Number
- ADA332675
Entities
People
- Dhan Kalvakolanu
- Edward Hofmann
Organizations
- University of Maryland, Baltimore