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Direct visualization of dispersed 11q13 chromosomal translocations in
mantle cell lymphoma by multicolor DNA fiber fluorescence in situ
hybridization
JW Vaandrager, E Schuuring, E Zwikstra, CJ de Boer, KK Kleiverda, JH van Krieken, HC Kluin-Nelemans, GJ van Ommen, AK Raap and PM Kluin
Department of Pathology, Leiden University Hospital, Leiden University, The
Netherlands.
Several hematologic malignancies are associated with specific chromosomal
translocations. Because of the dispersed distribution, chromosomal
breakpoints may be difficult to detect using molecular techniques. We
present a new application of a recently developed method, DNA fiber
fluorescence in situ hybridization (fiber FISH), which allows direct
visualization and mapping of chromosomal breakpoints. We tested this method
for detection of the t(11;14)(q13;q32) translocation in mantle cell
lymphoma. In DNA fiber FISH, a series of fluorochrome-labeled DNA probes
covering several hundreds of kilobasepairs is hybridized to linear DNA
molecules (or fibers) prepared from frozen tissue or intact cells. By using
alternate fluorescent colors, a potential breakpoint region is stained in a
color barcode pattern. Breaks in this region will split the barcode in two
complementary parts, from which the breakpoint position can be derived. We
used a 250-kb barcode covering the BCL-1 locus to detect 11q13 breakpoints
in 20 well-characterized mantle cell lymphomas. A t(11;14) was shown by
cohybridization of these probes with probes for the Ig heavy chain locus at
14q32. In 18 of 20 mantle cell lymphomas, a breakpoint within the
11q13/BCL-1 barcode was shown by the presence of multiple, complementary
translocation products. Fusion of 11q13 and 14q32 sequences on single
fibers indicating t(11;14)(q13;q32) was found in all 18 breakpoint-positive
mantle cell lymphomas. In one additional case, fusion of an intact 11q13
barcode with 14q32 sequences indicated a breakpoint 100 kb centromeric of
the major translocation cluster of BCL-1. Within the 120-kb region of
BCL-1, breakpoints were widely scattered. This explains why, so far, a
BCL-1 breakpoint had been detected by Southern blot analysis in only 10 of
19 cases. DNA fiber FISH analysis showed a t(11;14) in 95% of mantle cell
lymphoma. The results indicate that DNA fiber FISH is a rapid, simple, and
equally powerful method for detection of clustered and dispersed
translocation breakpoints.
Volume 88,
Issue 4,
pp. 1177-1182,
08/15/1996
Copyright © 1996 by The American Society of Hematology
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