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Application of Interphase Fluorescence In Situ Hybridization for the
Detection of the Burkitt Translocation t(8;14)(q24;q32) in B-Cell
Lymphomas
Reiner Siebert,
Peter Matthiesen,
Svetlana Harder,
Yanming Zhang,
Annekathrin Borowski,
Reina Zühlke-Jenisch,
Simone Metzke,
Stefan Joos,
Klaus Weber-Matthiesen,
Werner Grote, and
Brigitte Schlegelberger
From the Department of Human Genetics, University of Kiel, Kiel,
Germany; and Abteilung "Organisation komplexer Genome," Deutsches
Krebsforschungszentrum, Heidelberg, Germany.
The translocation t(8;14)(q24;q32) is the characteristic chromosomal
aberration of Burkitt's-type lymphomas and leukemias (BLs). On the
molecular level, the t(8;14) juxtaposes the c-myc gene in 8q24 next to
the IgH locus in 14q32, resulting in overexpression of the
transcription factor c-Myc. The detection of a t(8;14) is a major aim
in the diagnostic process of all patients with high-grade B-cell
lymphomas because treatment strategies differ between BL and other
high-grade lymphomas. As chromosome analyses are sometimes hampered by
the low yield or poor quality of metaphase spreads and as the
application of molecular genetic techniques is limited by the
distribution of the 8q24 breakpoints over a region of about some
hundred kilobases, we set out to establish an interphase fluorescence
in situ hybridization (FISH) assay for the detection of the t(8;14). A
cosmid probe hybridizing to the IgH constant region in 14q32 was
combined with a differently labeled probe of pooled cosmid clones
spanning the c-myc locus in 8q24. Interphase nuclei lacking a t(8;14)
show two separated signals corresponding to each probe, whereas
interphase nuclei carrying a t(8;14) display a split of the c-myc probe
and a colocalization of at least one of the splitted signals with the
IgH probe. Based on the results of extensive control studies, the
cutoff level for this stringent (type I) criteria was set at 2%.
Additionally, colocalization of at least one c-myc signal with one IgH
signal alone (without signal split for the c-myc probe) was used as a less stringent (type II) criteria with a cutoff limit of 11%. Nine BLs
and one Burkitt-like lymphoma were investigated by this approach.
Cytogenetically, all tumors contained a translocation t(8;14)(q24;q32)
except for one BL, in which cytogenetic analysis had failed. In
interphase FISH, all lymphomas and leukemias met the less stringent
criteria for the diagnosis of the t(8;14). Additionally, in all tumors
but the Burkitt-like lymphoma, a t(8;14) could be diagnosed according
to the stringent criteria. The percentage of cells found to harbor the
t(8;14) by FISH ranged from 4.3% to 100%. Comparison of cytogenetic
and FISH results revealed a significantly lower percentage of
t(8;14)+ interphase nuclei than metaphase cells
(P = .004). In conclusion, the described FISH assay
provides a feasible and sensitive tool for the routine detection of the
translocation t(8;14) in interphase cells which might also offer new
insights into the biology of high-grade B-cell lymphomas.
Blood, Vol. 91 No. 3 (February 1), 1998:
pp. 984-990
© 1998 by The American Society of Hematology.
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