Inhibition of Estrogen Receptor-Dependent Gene Transcription by a Designed Ligand.
Annual rept. 1 Jul 98-30 Jun 99
RESEARCH INST OF SCRIPPS CLINIC LA JOLLA CA
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The purpose of this study is to develop novel DNA ligands that offer the potential for the treatment of human breast cancer. Our approach is to develop small, cell-permeable molecules that prevent the activation of genes involved in breast cancer cell proliferation by the DNA-binding protein estrogen receptor. A series of pyrroleimidizole polyamides have synthesized in the laboratory of Dr. Peter Dervan at The California Institute of Technology and supplied to our laboratory. These polyamides were designed to bind the 6 bp half-site recognized by estrogen receptor. Standard DNase footprinting methods were used to measure the binding affinities of the synthetic ligands for their target sequences. A series of polyamides were screened for binding affinities and sequence specificity. We have used recombinant human estrogen receptor protein in DNA binding studies with the same target ERE sequences. Using DNase footprinting methods and gel mobility shift assays, we optimized conditions for ER-DNA interactions and we have shown that the ERE-binding polyamides inhibit ER binding to EREs. Future studies will examine whether these compounds are effective inhibitors of ER-dependent gene transcription in breast carcinoma cells in culture.
- Anatomy and Physiology
- Medicine and Medical Research