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

Abstract

Summary. Mantle-cell lymphoma (MCL) has a poorer prognosis than other small B-cell lymphomas, thus a definitive diagnosis is essential. The t(11;14)(q13;q32) associated with MCL juxtaposes portions of CCND1 (11q13) and IGH (14q32), resulting in over-expression of cyclin D1. In this study, a highly sensitive two-colour fluorescence in situ hybridization (FISH) method was developed to detect t(11;14)(q13;q32) in nuclei isolated from paraffin-embedded tissue. Twenty-three MCLs, 13 normal controls and nine small B-cell lymphomas other than MCL were studied by FISH. Each MCL had been previously investigated to detect genomic IGH±CCND1 fusion by polymerase chain reaction (PCR) using DNA extracted from frozen tissue. The IGH±CCND1 fusion detection rate in the MCLs was 96% by FISH compared with 35% by PCR. By FISH, one MCL and three small B-cell lymphomas other than MCL harboured abnormalities involving only IGH. Less than 1% of cells showed falsepositive IGH±CCND1 fusion in normal specimens by FISH. Thus, this highly sensitive FISH assay is very useful in confirming the diagnosis of MCL, has wide applicability as it may be performed on both paraffin-embedded and fresh tissue, and may also facilitate detection of translocations involving these loci in tumours other than MCL.

Immunohistochemical Detection of Cyclin D1 Using Optimized Conditions Is Highly Specific for Mantle Cell Lymphoma and Hairy Cell Leukemia

“…Our percentage of detection appears higher than that in other series; for example Ott cyclin D1 immunoreactivity in 88% of cases with proven t(11;14)(q13;q32), whereas Aguilera et al (16) found immunoreactivity in 70% of 23 cases with proven translocation. According to cytogenetic and Southern blotting analyses, approximately 50 to 80% of MCL show the t(11;14)(q13;q32) translocation (16 -18), and, according to fluorescence in situ hybridization, up to 96% show it (19,20). Thus, despite the small number of cases analyzed and the likelihood that in studying a substantive number of cases, the positivity rate may fall, it appears that our conditions for the immunohistochemical detection of cyclin D1 were highly sensitive, particularly in BMs.…”

“…Our results show that 6/10 cases of MCL had less than 25% of marrow involvement, that the neoplastic cells were cyclin D1 ϩ , and that none of the lymphocytes in reactive marrows showed cyclin D1 staining, in agreement with results from other studies (23,24). More recently, other approaches have been attempted to detect BM involvement; Remstein et al (19) and Caraway et al (25), in abstract forms, showed that fluorescence in situ hybridization is highly sensitive for detection of BM involvement by MCL, up to 96%; Caraway et al (25) showed that fluorescence in situ hybridization can detect t(11;14) breakpoints when polymerase chain reaction is negative.…”

A Comparative Analysis of FISH, RT-PCR, PCR, and Immunohistochemistry for the Diagnosis of Mantle Cell Lymphomas

“…Karyotype analysis allowed identification of t(11;14)(q13;q32) in only 70 to 75% of MCL, which may be related to the low mitotic index of malignant cells and to the poor morphology of metaphase spreads (9,27). Interphase fluorescence in situ hybridization (FISH) analysis circumvents these difficulties and has allowed the detection of t (11;14) in nearly 100% of MCL cases (8,28,29). Moreover, detection of t (11;14) by interphase FISH was achieved on cell suspensions or on nuclei isolated from fresh-frozen or paraffin-embedded material (8,29).…”

“…Interphase fluorescence in situ hybridization (FISH) analysis circumvents these difficulties and has allowed the detection of t (11;14) in nearly 100% of MCL cases (8,28,29). Moreover, detection of t (11;14) by interphase FISH was achieved on cell suspensions or on nuclei isolated from fresh-frozen or paraffin-embedded material (8,29). Competitive or semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR) allowed to detection of cyclin D1 transcript overexpression in nearly all MCL cases (7,16).…”

Real-Time RT-PCR Assay for Quantifying Cyclin D1 mRNA in B-Cell Non-Hodgkin's Lymphomas

“…More recently, fluorescence in situ hybridization methods have become popular because large probes can be used to detect the widely scattered breakpoints on chromosome 11. Using these methods, the t (11;14) can be detected in Ͼ95% of MCL cases (13)(14)(15).…”