In situ hybridization detection of cyclin Dl mRNA in centrocytic/mantle cell lymphoma

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Abstract
Background: Centrocytic/mantle cell lymphoma (MCL) is characterized by a specific chromosomal translocation, t(ll;14)(ql3;q32), which leads to deregulated expression of the Gl cyclin, cyclin Dl (PRADl, CCNDl, BCLl). Cyclin Dl overexpression has been demonstrated in MCL at the mRNA level by Northern blotting and at the protein level by both Western blotting and immunoperoxidase staining. Patients and methods: To assess the utility of in situ hybridization (ISH) to detect cyclin Dl mRNA expression in formalin-fixed, paraffin embedded tissue, five MCL specimens from three patients and two cases of B-cell smalllymphocytic lymphoma (B-SLL) were studied. BCL1 major translocation cluster gene rearrangements had been previously documented in two MCL patients; the other MCL and the two B-SLL showed no detectable BCL1 or cyclin Dl rearrangement. Results: ISH was performed using anti-sense 3H-labeled RNA probes for the cyclin Dl 3′ untranslated region (pPL7) and partial cyclin Dl cDNA (pPL8). ISH experiments usingan anti-sense actin RNA probe demonstrated adequate RNA preservation in all cases. Each of five specimens of MCL demonstrated increased cyclin Dl mRNA. In contrast, neither of the two cases of B-SLL demonstrated detectable levels. Conclusions: Overexpression of cyclin Dl mRNA can be detected in MCL by ISH using formalin fixed paraffin embedded tissue. The ISH technique may be useful in diagnosing and classifying low-grade B-cell lymphomas and should be applicable to the study of cyclin Dl mRNA expression in a broad spectrum of lymphoid proliferations and solid tumors.