Detection of Genetic Lesions in Breast Cancer
Abstract
The purpose of this proposal is to develop a "next generation' representational difference analysis protocol that uses transcribing nuclear DNA. A chromatin immunoprecipitation (ChIPs) technique is being established to isolate active chromatin by its association with highly acetylated hi stones. Before embarking with the ChIPs procedure, it was important to understand the dynamics of histone acetylation in breast cancer cells, and the effect of estradiol on these processes. In the past year, we determined the kinetics of histone acetylation in hormone dependent (T47D5) and hormone independent (MDA MB 231) breast cancer cells. For both cell lines, a small population (10%) of core histones is engaged in rapid hyperacetylation and rapid deacetylation. This population of core histones is thought to be associated with transcriptionally active DNA. The bulk of the acetylated core histones is slowly acetylated. We made the novel observation that estradiol increases the steady state level of histone acetylation in hormone dependent T47D5 breast cancer cells. Estradiol increased the steady state level of hi stone acetylation by decreasing the rate of histone deacetylation. The ChIPs protocol with anti-acetylated H3 antibodies was used to isolate transcriptionally active DNA from breast cancer cells.
Document Details
- Document Type
- Technical Report
- Publication Date
- Sep 01, 1998
- Accession Number
- ADA382537
Entities
People
- James Davie
Organizations
- University of Manitoba