Detection of the chromosomal translocation t(11;14) by polymerase chain reaction in mantle cell lymphomas

Abstract

The t(11;14)(q13;q32) and its molecular counterpart, BCL1 rearrangement, are consistent features of mantle cell lymphoma (MCL). Rearrangement is thought to deregulate the nearby CCND1 (BCL1/PRAD1) proto-oncogene, a member of the cyclin G1 gene family, and thereby to contribute to tumorigenesis. We and others have previously shown that the BCL1 locus is rearranged in 55% to 60% of MCL patients and that, on chromosome 11, more than 80% of the breakpoints are localized within a 1-kbp DNA segment known as the major translocation cluster (MTC). We have determined the nucleotide sequence for a portion of the MTC region, and constructed chromosome 11-specific oligonucleotides that were in conjunction with a consensus immunoglobulin (Ig) heavy chain joining region (JH) primer used to perform the polymerase chain reaction (PCR) to amplify t(11;14) chromosomal junctional sequences in DNA from 16 MCL patients with breakpoints in the MTC region. 15 of the 16 breakpoints that occurred at the MTC region were amenable to PCR detection. The sizes of the amplified bands, the existence or not of a Sac I site in the PCR products, and nucleotide sequencing of the amplified DNA from four patients showed that the breakpoints share a remarkable tendency to tightly cluster within 300 bp on chromosome 11, some of them occurring at the same nucleotide. On chromosome 14, the breakpoints were localized within the Ig JH. Our findings indicate that a BCL1 rearrangement can be detected using this approach in roughly one half of the MCL patients. This has implications for both the diagnosis and the clinical management of MCL.

Molecular analysis of bcl‐1/IgH junctional sequences in mantle cell lymphoma: potential mechanism of the t(11;14) chromosomal translocation

“…(B) Hybridization of the PCR amplification products corresponding to lanes 1-7 with a 32 P-dATP-labelled oligonucleotide probe (P4-11). Secondary bands of higher MW in lanes 4 and 5, that hybridize, represent PCR amplification products of bcl-1 with a J H 6 downstream of the J H gene (usually J H 4) that forms the junction, and observation also underscored by Rimokh et al (1994). probe (P4-11; 5 0 -TTTGGATAAAGGCGAGGAGC-3 0 ) internal to the primers used for the amplification reactions.…”

“…The identifiable D gene sequences were found to bear homology to DLR3, DM (Both DM1 and DM5) and DIR5 germline D genes (Table I). The analysis of the published junctional sequences yielded homology to germline D genes in 2/8 cases (Rimokh et al, 1994). Homology to germline D genes with a maximum of two base changes/deletions intervening was 10, 10 and nine nucleotides for samples 1406, 1413 and 3052, respectively (Table I).…”

“…As a result of this translocation, the bcl-1 proto-oncogene (11q13) is juxtaposed to the immunoglobulin (Ig) heavy chain junctional region (J H ) gene sequences (14q32) (Tsujimoto et al, 1984;Rimokh et al, 1993;Vandenberghe et al, 1992). The chromosomal translocation breakpoints are scattered over a large area on 11q13; however, the vast majority are known to cluster within a 1 kb segment of bcl-1 gene known as the major translocation cluster (MTC) (Tsujimoto et al, 1985;Rimokh et al, 1994).…”

Diagnostic utility of fluorescence in situ hybridization in mantle‐cell lymphoma

“…Cyclin D1 overexpression may be detected by paraffin section immunohistochemistry, but this is difficult both to accomplish and to interpret owing to the intense antigen retrieval necessary to optimize cyclin D1 immunoreactivity. The t(11;14)(q13;q32) has also been detected using polymerase chain reaction (PCR) and primers for a consensus immunoglobulin heavy chain joining region on chromosome 14 and a portion of the major translocation cluster (MTC) region on chromosome 11 (Williams et al, the wide variation in breakpoints along the cyclin D1 locus, the sensitivity of this technique ranges between only 27% and 57% (Rimokh et al, 1994;Luthra et al, 1995;Segal & Maiese, 1996;Chibbar et al, 1998).…”

Abnormal AP-1 protein expression in primary cutaneous B-cell lymphomas

“…34 Upregulation of CCND1 has been described in B-cell chronic lymphocytic leukaemia 35 and mantle cell lymphoma 36 as a consequence of a t(11;14)(q13;q32) translocation, leading to the juxtaposition of the CCND1 gene at 11q13 with the promoter region of the IgH gene at 14q32. 37,38 Increased CCND1 gene expression is also present in other B-cell lymphomas. [18][19][20] In this study, CCND1 protein was found to be expressed in a large proportion of PCBCL cases.…”