Regulation of Estrogen Receptor mRNA Stability in Human Breast Cancer Cells.
Abstract
The growth and metastases of approximately 40% of human breast cancers is dependent on 17beta-estradiol-estrogen receptor (E2-ER) complex. ER levels are controlled, in part, through an autoregulatory loop in which E2-ER complex (and mitogens) down-regulate ER mRNA levels - a process I have studied using Northern blot hybridization of RNAs extracted from MCF-7 human breast cancer cells. To resolve the ongoing controversy over the extent to which ER mRNA detabilization contributes to down-regulation, I had to develop new techniques for measuring mRNA degradation. I have created tetracycline regulated expression plasmids which transcribe either the full length 6.4 kb human ER mRNA or truncated mini-ER' mRNAs, identified improved methods for delivering the plasmids encoding these mRNAs to cells in transient transfections, and developed a novel quantitative PCR procedure, which for the first time in any system, allows quantitative PCR of RNA from cells transiently transfected with DNA plasmids encoding the RNA. Using this system, I will determine the specific sequences in ER mRNA important in estrogen and TPA destabilization and identify proteins binding to these sequences. This will provide important new information on mechanisms controlling ER mRNA levels in breast cancer cells.
Document Details
- Document Type
- Technical Report
- Publication Date
- Sep 01, 1997
- Accession Number
- ADA336697
Entities
People
- Vincent Dipippo
Organizations
- University of Illinois Urbana–Champaign