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RESEARCH PRODUCT
Detection of translocations affecting the BCL6 locus in B cell non-Hodgkin's lymphoma by interphase fluorescence in situ hybridization
Alessandro GozzettiHelen P. PriceDolors Sanchez-izquierdoJose A. Martinez-climentBrigitte SchlegelbergerI BenetM M Le BeauFrancesc SoléJavier García-condeLana HarderMartin J. S. DyerJesús M. Hernández-rivasReiner SiebertIsabel MarugánS GeskTakashi Sonokisubject
MaleCancer Researchmedicine.medical_specialtyLymphoma B-CellLymphomaMolecular Sequence DataTranslocationChromosomal translocationLocus (genetics)BiologyTranslocation GeneticFluorescenceChromosomesGeneticimmune system diseaseshemic and lymphatic diseasesmedicineHumansIn Situ Hybridization FluorescenceIn Situ HybridizationGeneticsGene Rearrangementmedicine.diagnostic_testBase SequenceBreakpointCytogeneticsB-CellBase Sequence; Chromosome Banding; Female; Gene Rearrangement; Humans; In Situ Hybridization Fluorescence; Karyotyping; Lymphoma B-Cell; Male; Molecular Sequence Data; Chromosomes Human Pair 3; Translocation GeneticHematologyGene rearrangementmedicine.diseaseMolecular biologyChromosome BandingOncologyChromosome 3KaryotypingPair 3FemaleChromosomes Human Pair 3TrisomyFluorescence in situ hybridizationHumandescription
Structural alterations in 3q27 affecting the BCL6 locus are among the most frequent changes in B-NHL. The aim of the present study was to establish an interphase-FISH assay for the detection of all diverse BCL6 translocations in B-NHL. Two different approaches were tested, one using a PAC-clone spanning the major breakpoint region (MBR) of BCL6 (span-assay), and another using two BAC clones flanking the MBR (flank-assay). Interphase FISH with the span-assay detected the various BCL6 translocations in seven B-NHL cell lines. The dual-color flank-assay was evaluated in two laboratories independently: in normal controls, the cutoff level for false-positive signals was 2.6%, whereas the cutoff level for false-negatives in the seven cell lines was 7.5%. To test the feasibility of the FISH strategies, 30 samples from patients with B-NHL with cytogenetic abnormalities of 3q27 were evaluated with both assays. In 21 cases, the span-assay indicated a BCL6 rearrangement. In 18 of the 21 cases, the dual-color flank-assay confirmed the translocation including 12 different partner chromosomal loci. The three false-positive cases detected with the span-assay showed trisomy of chromosome 3 by cytogenetic analyses, and they were correctly classified as non-rearranged with the flank-assay. In summary, our FISH strategy using two differently labeled flanking BCL6 BAC probes provides a robust, sensitive, and reproducible method for the detection of common and uncommon abnormalities of BCL6 gene in interphase nuclei. The routine application of this assay to patients with B-NHL will allow the assessment of the diagnostic and prognostic significance of BCL6 rearrangements.
| year | journal | country | edition | language |
|---|---|---|---|---|
| 2001-08-23 |