Expression Profiling of Cell Lines Expressing Regulated NP2 Transcripts
Abstract
Expression profiling is a powerful novel technique to examine changes in the expression of a large number of genes at the same time. Different phenotypic states of a cell can be translated into specific gene expression signatures. As a complement to yeast two hybrid studies we proposed using gene expression profiling to determine changes in gene expression as a function of expression of the neurofbromatosis-2 (NF2) gene in schwannoma cells. The strength of our approach is that we will not use tissues from patients, but will concentrate on cell lines in which NF2 expression can be controlled through the Tet/On system. In this system, treatment of cells with tetracycline (tet) induces expression of a tet-regulated gene, which in turn regulates the expression of the gene of interest. We have now generated several cell lines that express NF2 in a regulated fashion. The parent lines are RT4 schwannoma cells and mouse embryonic fibroblasts. A time course for NF2 expression has been established. A total of four cell lines have been tested on microarrays to detect expression changes. Surprisingly, no changes common to expression of isoform 1 and 2 have been detected so far. In the last year, we have used cell lines that express HRS in the tet system to study expression changes induced by HRS in the presence or absence of EGF. We established a time-course of EGF-induced genes and then examined the effects of long-term treatment (18hrs) with EGF in the presence or absence of exogenous HRS. The results will provide a framework for the interpretation of future gene expression studies in vitro and in NF2 tumors.
Document Details
- Document Type
- Technical Report
- Publication Date
- Sep 01, 2004
- Accession Number
- ADA436907
Entities
People
- Stefan M. Pulst
Organizations
- Cedars-Sinai Medical Center