Detection of three common translocation breakpoints in non‐Hodgkin's lymphomas by fluorescence in situ hybridization on routine paraffin‐embedded tissue sections

Abstract

Non-random chromosomal translocations are specifically involved in the pathogenesis of many non-Hodgkin's lymphomas and have clinical implications as diagnostic and/or prognostic markers. Their detection is often impaired by technical problems, including the distribution of the breakpoints over large genomic areas. This study reports a fluorescence in situ hybridization (FISH) method which allows the detection of specific chromosomal breakpoints in tissue sections from routinely fixed, paraffin-embedded samples. Hybridization was performed after demasking the DNA. Previously validated locus-specific probes (cosmids, PACs) flanking the BCL1, BCL2 regions and similar new probes for the MYC breakpoint region were used. The cases studied were five mantle cell lymphomas (MCL) and five follicular lymphomas (FL), selected on the basis of a previously proved t(11;14) and t(14;18) and five randomly chosen Burkitt's lymphomas (BL), as well as 21 negative control samples. In all samples, hybridization signals of sufficient intensity were obtained. Three different algorithms were used to score the hybridization signals in tissue sections, two of them taking into account the nuclei and their signal distribution indicative of chromosomal break, and one only considering the colocalization or segregation of the signals. In control tissues, these algorithms resulted in cut-off levels of 9.1%, 1.3%, or 10.0%. In the 15 lymphoma samples the percentages of abnormal cells/signals ranged from 28% to 80%, 13% to 49%, and 40% to 70%, respectively. The results indicate that small locus-specific probes can be used in FISH for regular detection of translocation breakpoints on routine paraffin tissue sections.

Exclusion of RUNX3 as a tumour-suppressor gene in early-onset gastric carcinomas

“…Labelling, hybridization and detection were performed as described previously (Haralambieva et al, 2002), with the exception of the final probe concentration (30 ng/ml). The probes used were PAC LLNLP704N11132 (RZPD, Berlin, Germany) (RUNX3) and pUC1.77 (centromere 1).…”

“…Pretreatment of Y4 paraffin sections was performed as described previously (Haralambieva et al, 2002).…”

Design and standardization of PCR primers and protocols for detection of clonal immunoglobulin and T-cell receptor gene recombinations in suspect lymphoproliferations: Report of the BIOMED-2 Concerted Action BMH4-CT98-3936

“…2 The gold standard detection strategy for the presence of the t(11;14) that will identify almost all breakpoints is interphase FISH using breakpoint-flanking probes in fresh or frozen material 140 as well as in archival specimens. 144 However, a PCR-based detection strategy for the t(11;14) might be useful for residual disease monitoring. Many groups have developed PCR-based assays to detect the BCL1/JH breakpoints, in general using a consensus JH primer in combination with primers in the BCL1-MTC region that were all located in a region of 392 bp.…”

Double-staining chromogenic in situ hybridization as a useful alternative to split-signal fluorescence in situ hybridization in lymphoma diagnostics

“…It is known that different types of lymphoma are associated with non-random chromosomal translocations. 1,2,3,4,5 The detection of these aberrations is, therefore, an important step in the identification of specific lymphoma entities. Several techniques are now available to detect chromosomal alterations.…”