Detection of the Chromosomal Translocation t(l1; 14) by Polymerase Chain Reaction in Mantle Cell Lymphomas zyx By Ruth Rimokh, FranGoise Berger, Georges Delsol, Isabelle Digonnet, Jean Pierre Rouault, Jean Dominique Tigaud, Mylene Gadoux, Bertrand Coiffier, Paul Andre Bryon, and Jean Pierre Magaud The t(11;14)(q13;q32) and its molecular counterpart, zyxwvut BCLl rearrangement, are consistent features of mantle cell lymphoma (MCL). Rearrangement is thought to deregulate the nearby CCNDl (BCLl/PRADl) proto-oncogene, a mem- ber of the cyclin zyxwvutsrq G, gene family, and thereby to contribute to tumorogenesis. We and others have previously shown that the BCLl locus is rearranged in 55% t o 60% of MCL patients and that, on chromosome 11, more than 80% of the breakpoints are localized within a l-kbp DNA segment known as the major translocation cluster (MTCI. We have determined the nuclaotide sequencefor a portion of the MTC region, and constructed chromosome 1 l-specific oligonucle- otides that were in conjunction with a consensus immuno- globulin (Ig) heavy chain joining region (JH) primer used to perform the polymerasechainreaction (PCR) to amplify zyxwvuts ALIGNANT NON-HODGKIN'S lymphomas and many other human tumors are often associated with specific chromosome translocations, and experimental evi- dence indicates that genes located at recumng chromosomal breakpoints are directly involved in tumor pathogenesis.' Furthermore, several of these cytogenetic abnormalities are potentially valuable as diagnostic and prognostic aids. The most frequent translocation in human lymphoma is the t(14;18)(q32;q21)? which can be detected by cytogenetic or molecular analyses in approximately 85% of follicular lymphomas. This translocation juxtaposes the proto-onco- gene BCL2, from 18q21, with an immunoglobulin (Ig) heavy chain joining region (JH) segment on the derivative chromo- some 14. This results in a BCL2-Ig fusion gene, chimeric transcripts, and transcriptional deregulation of the translo- cated BCL2.3.4 The chromosome breakpoints are remarkably well focused on chromosome segment 18q21 in two well- defined regions of 500 bp, known as the major and the minor breakpoint regi0ns.4.~ Taking advantage of this situation, sev- eral authors have shown that the t(14; 18) can be detected by polymerase chain reaction (PCR) amplification of DNA extracted from fresh tumor cells, frozen tissues, and even from formalin-fixed tissue^.^.^ Moreover, because of its ex- tremely high sensitivity, PCR amplification of the BCL2-Ig junction offers a valuable means of detecting minimal resid- ual disease in follicular non-Hodgkin's lymphomas. Mantle cell lymphoma (MCL) represents a clinicopatho- logically distinct subtype of malignant non-Hodgkin's lymphomas that originates from immature virgin B cell in the primary follicle and mantle zone of peripheral lymphoid It accounts for 5% to 10% of the overall number of malignant non-Hodgkin's lymphomas in adults. The t(l1; 14)(q13;q32) and its molecular counterpart, BCLI re- arrangement, are consistent features of this lymphoma sub- type.'"I6 As a result of the t(l1; 14), the BCLl locus is juxta- posed to an Ig enhancer sequence located on chromosome 14.'7*'8 The subsequent deregulation of zyxwvutsr CCNDl (PRADU BCLl), a member of the cyclin GI gene family, is thought to perturb the GI-S transition of the cell cycle and thereby to contribute to tumor de~elopment.l~-~~ Although the chro- M Blood, Vol zyxwvutsrqpon 83, No 7 (April I), 1994 pp 1871-1875 t(11;14) chromosomal junctional sequences in DNA from 16 MCL patients withbreakpoints in the MTC region. 15 of the 16 breakpoints that occurred at the MTC region were amena- ble t o 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 pa- tients showed that the breakpoints share a remarkable ten- dency to tightly cluster within 300 bp on chromosome 11, some of them occurring at the same nucleotide. On chromo- some 14, the breakpoints were localized within the Ig JH. Our findings indicate that a BCLl rearrangement can be de- tected using this approach in roughly one half of the MCL patients. This has implications for both the diagnosis and the clinical management of MCL. zyx 0 1994 b y The American Society of Hematology. mosome breakpoints are widely scattered on chromosome region 1 lq13, weand others have shown thatmorethan 80% of them are clustered in a restricted DNA segment known as the major translocation cluster (MTC).'3.'5"8.24 In the present study we showed that 15 of the 16 t(l1; 14) breakpoints that occur at the MTC are amenable toPCR detection because they share a remarkable tendency to clus- ter tightly within 300 bp on chromosome 1 1. These findings should facilitate application of the PCR techniques for the diagnosis and the clinical management of MCL. MATERIALS AND METHODS Tumoral samples and cell lines. Characterization of the tumoral samples and cell lines used in this study have been previously re- ported (cases 1 to 39)? All the MCL cases analyzed here showed a prominent diffuse pattern of growth. The human B-cell line Rec- 1 (referred to as case 40) was established from a t(l1; 14)(q13;q32)- bearing lymphoma.2' DNA probes and hybridization procedures. The probe for the Ig JH was a gift from T.H. Rabhitts (MRC, Cambridge, UK). The MTC probe was kindly given to us by Y. Tsujimoto (Wistar Institute, Philadelphia, PA) and consists of a 2.3-kbp Sac I-Sac I DNA geno- mic fragment. The EH04 probe corresponds toan EcoRI-Hind111 From the Laboratoire d'Himatologie et de Cytoginitique and the Laboratoire d'Anatomie pathologique, HBpital Edouard Herriot; and the Dipartement d'Himatologie, Centre Hospitalier Lyon Sud, Lyon, France. Submitted September 22, 1993; accepted November 17, 1993. Supported by grants from INSERM (CRE 92-0108, CJF 93-07), CNRS, Hospices Civils de Lyon, and Ligue Nationale Contre le Cancer (Comiti du RhBne, de 1'Yonne et de la SaBne et hire). Address reprint requests to Ruth Rimokh, PhD, Laboratoire d'Him- atologie et de Cytogknktique, Pavillon E, HBpital Edouard Herriot, 69437 Lyon Cedex 03, France. The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. section 1734 solely to indicate this fact. zyxwv 0 1994 by The American Society of Hematology. 0006-4971/94/8307-0002$3.00/0 1871 Downloaded from http://ashpublications.org/blood/article-pdf/83/7/1871/613972/1871.pdf by guest on 21 January 2023