Three-color FISH analysis of TMPRSS2/ERG fusions in prostate cancer indicates that genomic microdeletion of chromosome 21 is associated with rearrangement - PubMed (original) (raw)
Three-color FISH analysis of TMPRSS2/ERG fusions in prostate cancer indicates that genomic microdeletion of chromosome 21 is associated with rearrangement
Maisa Yoshimoto et al. Neoplasia. 2006 Jun.
Abstract
The recent description of novel recurrent gene fusions in approximately 80% of prostate cancer (PCa) cases has generated increased interest in the search for new translocations in other epithelial cancers and emphasizes the importance of understanding the origins and biologic implications of these genomic rearrangements. Analysis of 15 PCa cases by reverse transcription-polymerase chain reaction was used to detect six ERG-related gene fusion transcripts with TMPRSS2. No TMPRSS2/ETV1 chimeric fusion was detected in this series. Three-color fluorescence in situ hybridization confirms that TMPRSS2/ERG fusion may be accompanied by a small hemizygous sequence deletion on chromosome 21 between ERG and TMPRSS2 genes. Analysis of genomic architecture in the region of genomic rearrangement suggests that tracts of microhomology could facilitate TMPRSS2/ERG fusion events.
Figures
Figure 1
Rearrangement of the TMPRSS2 and ERG genes in PCa. (A) RT-PCR products from six PCa cases were sized using the Agilent 2100 Bioanalyzer. The fragments were analyzed with a ladder marker to determine the size of each variant TMPRSS2/ERG transcript. Depending on the breakpoints within each, the fragments were 800, 600, ∼430, and ∼350 bp. (B) Sequence electropherograms of mutant TMPRSS2/ERG transcripts from case 78-01. Two unique variant transcripts were found to be present in this case: one containing exons 1and 2 of the TMPRSS2 gene and exons 5 and 6 of the ERG gene, and the other containing exon 1 of the TMPRSS2 gene joined to exons 5 and 6 of the ERG gene. The arrows indicate gene breakpoints. (C) Schematic representation of the exon composition of the TMPRSS2/ERG gene fusion products from variant PCa cases.
Figure 2
Location and names of the BAC probes and gene locations used in the analysis. Gene locations are taken from the May 2004 assembly of the UCSC Genome Browser. Numbers indicate basepair location along the chromosome. Colors correspond to fluorochromes used in FISH experiments.
Figure 3
FISH analysis showing rearrangement of TMPRSS2 and ERG genes in PCa. (A) FISH confirms the colocalization of OregonGreen-labeled 5′ ERG (green signals), AlexaFluor 594-labeled 3′ ERG (red signals), and Pacific Blue-labeled TMPRSS2 (light blue signals) in normal peripheral lymphocyte metaphase cells and in normal interphase cells. (B) In PCa cells, break-apart FISH results in a split of the colocalized 5′ green/3′ red signals, in addition to a fused signal (comprising green, red, and blue signals) of the unaffected chromosome 21. Using the TMPRSS2/ERG set of probes on PCa frozen sections, TMPRSS2 (blue signal) remains juxtaposed to ERG 3′ (red signal; see white arrows), whereas colocalized 5′ ERG signal (green) is lost, indicating the presence of TMPRSS2/ERG fusion and concomitant deletion of 5′ ERG region.
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