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Published on: October 8, 2015
Molecular diagnosis of mantle cell lymphoma in paraffin-embedded tissue
J Lasota1, K Franssila, C H Koo
1Department of Pathology, Anatomy and Cell Biology, Jefferson Medical College of Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA.
Insights
Mantle cell lymphoma diagnosis can be aided by a new nested polymerase chain reaction (PCR) test detecting the t(11;14) chromosomal translocation. This PCR method shows promise for identifying mantle cell lymphoma in fixed tissues, though its diagnostic power is limited.
Area of Science:
- Hematology
- Oncology
- Molecular Biology
- Genetics
Background:
- Mantle cell lymphoma (MCL) is an aggressive non-Hodgkin lymphoma requiring differentiation from other small B-cell lymphomas.
- MCL is characterized by the t(11;14) chromosomal translocation, involving the bcl-1 oncogene and immunoglobulin heavy chain gene.
Purpose of the Study:
- To develop and evaluate a nested polymerase chain reaction (PCR) system for detecting the t(11;14) translocation in MCL.
- To assess the utility of this PCR method in identifying MCL, particularly in fixed tissue samples.
Main Methods:
- A novel nested PCR system was designed to amplify bcl-1 breaks within the major translocation cluster.
- The system involved two successive PCR amplifications, followed by Southern blotting and restriction enzyme digestion for product confirmation.
- The method was tested on 10 MCL cases, other lymphomas, and hyperplastic lymph nodes.
Main Results:
- The nested PCR system successfully generated a specific product in 4 out of 10 MCL cases.
- Southern blotting and restriction enzyme analysis confirmed the identity and variability of the amplified products.
- All other tested lymphoma types and hyperplastic lymph nodes were negative for the translocation.
Conclusions:
- The nested PCR system provides a convenient method for identifying the t(11;14) translocation in MCL, especially in formaldehyde-fixed, paraffin-embedded tissues.
- The test has limited diagnostic utility as only about half of MCL cases exhibit bcl-1 breaks in the major translocation cluster.
- This technique facilitates retrospective studies of MCL using archival tissue samples.
Abstract:
Mantle cell lymphoma (formerly known as intermediate lymphocytic lymphoma, diffuse centrocytic lymphoma, or diffuse small cleaved lymphoma) is one of the small cell non-Hodgkin lymphoma entities that is clinically more aggressive than small lymphocytic lymphoma, and needs to be separated from it. Mantle cell lymphoma is strongly associated with the t(11;14) chromosomal translocation that rearranges the bcl-1 oncogene (PRAD-1 gene) and immunoglobulin heavy chain gene. In this study, we developed a nested polymerase chain reaction system to evaluate the t(11;14) translocation. The study material consisted of 10 mantle cell lymphomas fulfilling the criteria suggested by other authors (P. M. Banks et al. Surg Pathol 16:637, 1992). A novel nested polymerase chain reaction system was used to evaluate the bcl-1 breaks in the major translocation cluster using two successive polymerase chain reaction amplifications. This reaction yielded a background-free single product of the size of 200 to 300 base pairs in four of 10 mantle cell lymphomas. The identity of the product from the nested polymerase chain reaction was confirmed by Southern blotting followed by hybridization with a specific probe. The amplification products were also evaluated by Sst-I, Alu-I, Dde-I, and Ita-I restriction enzymes and showed different patterns of digestion reflecting individual differences between the MTC/ IgH junctions. A selection of other low-grade lymphomas, including lymphocytic, follicular, and mucosa-associated lymphoid tissue lymphoma, and hairy cell leukemia and 29 hyperplastic lymph nodes were negative. This nested polymerase chain reaction system for the t(11;14) translocation involving major translocation cluster offers a convenient specific identification for mantle cell lymphoma. However, this test has a limited diagnostic power because only about half of the mantle cell lymphomas show the bcl-1 breaks in the major translocation cluster. The test performs well in formaldehyde-fixed and paraffin-embedded material, allowing the study of large numbers of retrospective cases of mantle cell lymphomas.
