Identification of Prostate Cancer-Related Genes Using Inhibition of NMD in Prostate Cancer Cell Lines
Abstract
A strategy to identify mutant genes using inhibition of nonsense mediated decay (NMD) in cell lines has been proposed by others. Blocking translation with antibiotic emetine has been shown to inhibit the NMD. Stabilization of mutant mRNA following the inhibition of NMD with emetine can be detected using microarray technology, such as Affymetrix genechips, for example. Unfortunately, too many genes that do not contain any mutations show mRNA increase following emetine treatment due to stress response to the inhibition of translation or due to being a natural substrate for NMD, thus complicating the identification of mutant genes. We have developed a simple analytical method that increases the efficiency of identifying mutant genes in cell lines following inhibition of nonsense mediated decay (NMD). The approach assumes that the spectra of mutant genes differ between cell lines of the same tumor origin. Using this analytical filter we have identified previously unknown inactivating mutations in the Jak1, Synaptojanin2 (SYNJ2) and Cleft Lip and Palate Transmembrane Protein 1(CLPTM1) genes in the prostate cancer cell line LANCaP. A list of candidate genes for sequencing analysis has been generated for the other prostate cancer cell lines.
Document Details
- Document Type
- Technical Report
- Publication Date
- Jan 01, 2005
- Accession Number
- ADA435274
Entities
People
- Yurij Ionov
Organizations
- Health Research, Incorporated