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 screened siRNA designed in our lab or commercially available siRNA targeting each FGFR and shown their ability to reduce target mRNA levels using transient transfection assays. These assays have indicated that siRNAs designed against each receptor are able to at least partially reduce target mRNA as determined by quantitative RT-PCR. Using these sequences, we have developed clonal and polyclonal cell lines that stably express shRNA against all 4 FGFRs. We have also developed shRNA expressing vectors that also express EGFP and are currently using these vectors to determine cell cycle progression in ML-20 cells expressing the shRNA under FGF dependent conditions.
Document Details
- Document Type
- Technical Report
- Publication Date
- May 01, 2004
- Accession Number
- ADA425972
Entities
People
- Norman R. Estes
Organizations
- Southern Research