INVOLVEMENT OF THE CCND1 GENE IN HAIRY-CELL LEUKEMIA

Background: Previous results suggested increased mRNA expression of CC ND1 in hairy cell leukemia (HCL). The CCND1 gene is involved in the t( 11;14)(q13;q32) chromosomal rearrangement, a characteristic abnormalit y in mantle cell lymphoma (MCL). We and others reported that, in contr ast to other B-cell lymphomas, almost all MCL have overexpression of t he CCND1 gene with a good correlation between RNA and protein analysis . Recent studies showed that overexpression of the cyclin D1 protein c an be easily detected by immunohistochemistry (IHC) on formalin-fixed, paraffin embedded tissues. Patients and methods: To investigate wheth er the CCND1 gene is involved in HCL, we performed IHC on a series of 22 cases using formalin-fixed paraffin embedded splenectomy specimens. For IHC the sections were boiled in citrate buffer. The presence of r earrangements within the BCL-1 locus and the CCND1 gene was analyzed i n 13 of 22 cases by Southern blot analysis using all available breakpo int probes. Expression of CCND1 was analyzed at the mRNA level (Northe rn blot) and protein level (IHC). Results: Overexpression of the cycli n D1 protein using IHC was observed in all cases, with strong expressi on in 5 cases. Pre-existing B- and T-cell areas of the spleen did not express significant levels of the cyclin D1 protein. Seven of 9 cases analyzed by both IHC and Northern blotting showed overexpression of th e CCND1 gene with both methods. No genomic abnormalities were observed in any of the 13 cases studied by Southern blot analysis. Additionall y, no 11q13 abnormalities were detected by banding analysis of 19 of 2 2 cases. Conclusions: The elevated levels of CCND1 mRNA and protein in conjunction with the absence of overt rearrangements within the BCL-1 locus distinguish HCL from MCL and other B-cell malignancies. This su ggests that activation of the CCND1 gene in HCL is due to mechanisms o ther than chromosomal rearrangement.