Modulation of the Proliferation and Metastasis of Human Breast Tumor Cells by SLUG (IDEA)
Abstract
The objective of the project for the reporting period was to identify high affinity SLUG-regulated gene promoters from human breast cells. We over expressed 3xFLAG-tagged (C-terminal) human SLUG in the SLUG-negative MDA-MB-468 and MCF-7 cells through a lentiviral construct. By chromatin immunoprecipitation assay and Q-PCR, we identified/reconfirmed a handful of the gene promoters that indeed bind in vivo with SLUG in the human breast cells tested. They include cytokeratins 8, 18, and 19, E-cadherin, occludin, Na/K ATPase, vitamin D receptor, integrin alpha 3, PUMA, BRCA2, claudins 5, 3, 7, 11, 1, 14, and 16, desmoglein 1, and 2, PCNA, PGDH and plakoglobin gene promoters. We are employing the chromatin immuno-precipitation-DNA selection and ligation (ChIP-DSL, formerly known as ChIP-GLAS, Aviva System Biology) technique for further analysis. We evaluated the mRNA levels of these potentially SLUG-regulated genes in invasive and non-invasive human breast tumor cells. We have designed DNA decoys against the DNA binding domain of SLUG from the claudin 7 gene promoter (highest affinity for SLUG among the promoters tested). We are currently testing whether these decoys can block the binding of SLUG in vivo to its target promoters and stimulate the expressions of SLUG-regulated genes in human breast cells.
Document Details
- Document Type
- Technical Report
- Publication Date
- Apr 01, 2007
- Accession Number
- ADA472059
Entities
People
- Gautam Chaudhuri
Organizations
- Meharry Medical College