Mantle cell lymphoma: improved diagnostics using a combined approach of immunohistochemistry and identification of t(11;14)(q13;q32) by polymerase chain reaction and fluorescence in situ hybridization.
BACKGROUND
Mantle cell lymphoma (MCL) is a clinicopathological entity characterized by an aggressive clinical course, morphological features, and overexpression of cyclin D1 due to juxtaposition of the bcl-1 locus (and CCND1 gene coding for the cyclin D1) to the IgH gene. This phenomenon is caused by t(11;14)(q13;q32). The morphological diagnosis of MCL may pose difficulties. Ancillary methods are available to support the diagnosis.


METHODS
We studied a group of 32 patients with MCL; 24 men and 8 women. The median age at the diagnosis was 64 years. We characterized the investigated group by histology, and to analyze the immunohistochemical (IHC) profile we used a panel of antibodies including anti-cyclin D1. Polymerase chain reaction (PCR) was used to detect the rearrangement of bcl-1/IgH in 26 cases (in 11 patients, the DNA was isolated from frozen tissues or from nucleated cells of bone-marrow aspirate or peripheral blood, in 15 patients we utilized paraffin-embedded material). Dual color fluorescence in situ hybridization (FISH) on interphase nuclei detecting the t(11;14)(q13;q32) was applied in all 32 cases.


RESULTS
Cyclin D1 IHC was positive in 29 of 30 cases tested (97%). In six, the result was weak and difficult to rely on to support the diagnosis. PCR revealed the fusion gene in 14 of the 26 cases (54%). The best yield was obtained from fresh and frozen samples (8 of 11 positive). Using FISH, we identified the translocation in all 32 patients, the findings being easily interpretable in 29 patients. In three cases, the intensity of red and green signals was weaker and difficult to read though the co-hybridized signals were identified. The classical pattern of the translocation was observed in 26 patients, while in 3 we found variant patterns suggesting a loss of the V segment of the IgH gene (2x) and a shift in the breakpoint region at chromosome 11 (1x).


CONCLUSIONS
The diagnosis of MCL should be supported by a complex laboratory approach. Interphase FISH seems a useful complementary method to morphology and IHC. It is applicable to various tissues and cells prepared as tissue imprints or histological sections.
Kodet
Roman
R
Department of Pathology and Molecular Medicine, Charles University, 2nd Medical School and Faculty Hospital in Motol, V Uvalu 84, 150 06, Prague 5-Motol, Czech Republic. Roman.Kodet@lfmotol.cuni.cz
Mrhalová
Marcela
M
Krsková
Lenka
L
Soukup
Jan
J
Campr
Vít
V
Neskudla
Tomás
T
Szépe
Peter
P
Plank
Lukás
L
D000368:Aged; D000369:Aged, 80 and over; D014408:Biomarkers, Tumor / Q000032:analysis; D002880:Chromosomes, Human, Pair 11; D002883:Chromosomes, Human, Pair 14; D019938:Cyclin D1 / Q000032:analysis; D004273:DNA, Neoplasm / Q000032:analysis; D005260:Female; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D020522:Lymphoma, Mantle-Cell / Q000737:chemistry / Q000175:diagnosis* / Q000235:genetics; D008297:Male; D008875:Middle Aged; D016133:Polymerase Chain Reaction; D012189:Retrospective Studies; D014178:Translocation, Genetic
D014408:Biomarkers, Tumor; D004273:DNA, Neoplasm; D019938:Cyclin D1