Analysis of the transcriptional activity of amplified genes in tumour cells by fluorescence in situ hybridization.
In this paper we present a new application of the detection of nuclear transcripts by fluorescence in situ hybridization (FISH) for studying the transcriptional activity of amplified genes in tumour cells. As a model, we have used the A431 cell line in which several amplification sites have been identified. We focused on two amplified regions: (1) the 6p12 region, which was found amplified by using comparative genomic hybridization, and which contains an amplification of the hsp90 beta gene; (2) the 7p12-p13 region, which displays a 20- to 30-fold amplification of the gene encoding the epidermal growth factor receptor (EGFr). By using FISH to detect nuclear transcripts, we show that the extra-copies of the hsp90 beta and EGFr genes are actively transcribed within the sites of amplification. This work illustrates the potential of this method as a tool for functional in situ cytogenetic analyses.
Jolly
C
C
Inserm U309, Equipe DyOGen, Institut Albert Bonniot, Université Joseph Fourier Grenoble I, La Tronche, France. caroline.jolly@ujf-grenoble.fr
Michelland
S
S
Rocchi
M
M
Robert-Nicoud
M
M
Vourc'h
C
C
D002874:Chromosome Mapping; D002896:Chromosomes, Human, Pair 6 / Q000235:genetics; D002897:Chromosomes, Human, Pair 7 / Q000235:genetics; D066246:ErbB Receptors / Q000235:genetics; D005784:Gene Amplification; D015321:Gene Rearrangement; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics; D018841:HSP90 Heat-Shock Proteins / Q000235:genetics; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D012878:Skin Neoplasms / Q000235:genetics*; D014158:Transcription, Genetic; D014407:Tumor Cells, Cultured
D018840:HSP70 Heat-Shock Proteins; D018841:HSP90 Heat-Shock Proteins; D066246:ErbB Receptors