bcl-1 gene rearrangements in mantle cell lymphoma: a comprehensive analysis of 118 cases, including B-5-fixed tissue, by polymerase chain reaction and Southern transfer analysis.
We evaluated 118 cases of mantle cell lymphoma by polymerase chain reaction (PCR) for the major translocation cluster (MTC) region and another breakpoint corresponding to probe p94PS, located 24 kb telomeric to the MTC locus on chromosome 11. The specimens included 64 frozen, 19 formalin-fixed, and 9 B-5-fixed lymph nodes and 26 B-5-fixed bone marrow biopsy specimens. We also analyzed DNA from the 64 frozen lymph nodes by Southern transfer analysis (SB) using three separate bcl-1 breakpoint probes. Gene rearrangements were identified in 17 (PCR) and 18 (SB) of 64 frozen lymph nodes and by PCR in 6 of 19 formalin-fixed lymph nodes, 3 of 9 B-5-fixed lymph nodes, and 12 of 26 B-5-fixed bone marrow cores with MTC locus primers and probe. Only one case showed rearrangement with the p11EH probe that corresponds to breakpoints situated 63 kb telomeric to the MTC locus. No rearrangements were detected by PCR or SB for the breakpoint site corresponding to the p94PS probe, but we identified a polymorphic restriction site with HinD III digest in approximately 25% of the cases. In agreement with other studies, these results confirmed that breakpoints in the MTC region of the bcl-1 locus are tightly clustered and associated with 30 to 40% of mantle cell lymphomas. Other breakpoints in the bcl-1 locus seem to be heterogeneous and cannot be detected by PCR or SB with use of existing probes or primer sequences. The most important finding of our study is optimization of the methodology for the detection of immunoglobulin heavy chain gene rearrangement and MTC region breakpoints by PCR from the DNA isolated from B-5-fixed, paraffin-embedded lymph nodes and bone marrow biopsy specimens. The results obtained from these tissues are comparable to those obtained from frozen or formalin-fixed tissue.
Chibbar
R
R
Department of Laboratory Medicine and Pathology, University of Minnesota, Minneapolis, USA. rchibbar@gpu.srv.ualberta.ca
Leung
K
K
McCormick
S
S
Ritzkalla
K
K
Strickler
J
J
Staggs
R
R
Poppema
S
S
Brunning
R D
RD
McGlennen
R C
RC
D015139:Blotting, Southern; D002873:Chromosome Fragility; D002999:Clone Cells; D004247:DNA / Q000032:analysis / Q000235:genetics; D015342:DNA Probes; D004273:DNA, Neoplasm / Q000032:analysis / Q000235:genetics; D005404:Fixatives; D015321:Gene Rearrangement; D019940:Genes, bcl-1 / Q000235:genetics*; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000235:genetics; D008228:Lymphoma, Non-Hodgkin / Q000235:genetics*; D016133:Polymerase Chain Reaction
D015342:DNA Probes; D004273:DNA, Neoplasm; D005404:Fixatives; D007143:Immunoglobulin Heavy Chains; D004247:DNA