Hormonal Involvement in Breast Cancer Gene Amplification
Abstract
We propose to map origins of replication in the breast cancer genome and compare these sites with genomic positions that bind the estrogen receptor (ER) and genomic regions of DNA amplification. Correlations will support our hypothesis that ER adjacent to replication origins may interact with the replication machinery to drive DNA amplification which is a hallmark of many cancers. To work out the methodology, we have begun our experiments with thwell-studied breast cancer cell line MCF7 grown by the Brodsky lab. The Gerbi lab has refine the lambda exonuclease method to obtain nascent DNA strands (up to 20 times enriched) and hashown that the myc origin maps to the same map position in HeLa and MCF7 cells. Soon we will use the nascent DNA (size fractionated to 500-1500 nt) for next generation sequencing which gives better resolution and greater sensitivity than DNA microarrays. The Brodsky lab is testing the use of ORC antibodies for chromatin immunoprecipitation (ChIP) which will validate our replication origin map based on nascent strands.
Document Details
- Document Type
- Technical Report
- Publication Date
- Oct 01, 2008
- Accession Number
- ADA501716
Entities
People
- Alexander Brodsky
- Ben Raphael
- Susan A. Gerbi
Organizations
- Brown University