Inactivation of FGF Receptors by Targeting Ribozymes Against FGFR mRNAs and Their Effect on FGF Dependent In Vitro and In Vivo Breast Cancer Growth Phenotypes
Abstract
Acquired tamoxifen resistance is common in breast cancer patients with estrogen receptor positive (ER+) tumors. Growth factor signaling can provide ER+ breast cancer cells with alternative growth stimulus to that provided by activation of ER. In order to determine whether an individual FGF receptor (FGFR) or multiple receptors are responsible for conferring an alternate growth signaling pathway, we are using a siRNA targeting strategy to selectively inactivate each of the receptors either singly or in combination. We have designed multiple siRNA against each of the four receptors and screened their ability to reduce target mRNA levels using transient transfection assays. These assays have indicated that at least 3 of 4 individual siRNA against each receptor is able to at least partially reduce target mRNA as determined by quantitative RT-PCR. Using one of these siRNA sequences, we have developed clonal and polyclonal cell lines that stably express siRNA against EGFR3 and shown that these are able to reduce target mRNA up to 90%. We are currently screening additional siRNA against the remaining receptors. These results indicate the feasibility of designing siRNA against the FGFRs and developing stable clones that are able to efficiently knock down target mRNA.
Document Details
- Document Type
- Technical Report
- Publication Date
- May 01, 2003
- Accession Number
- ADA416690
Entities
People
- Francis G. Kern
- Norman R. Estes
Organizations
- Southern Research