Quantitative real-time reverse transcription-PCR assay for cyclin D1 expression: Utility in the diagnosis of mantle cell lymphoma

Background: The t(11;14)(q13;q32) translocation present in the majority of mantle cell lymphomas (MCLs) places the cyclin DI gene under the control of immunoglobulin transcriptional regulatory elements, causing overexpression of cyclin D1. Quantification of cyclin D1 expression can distinguish MCL f rom other lymphomas. Methods: A quantitative real-time reverse transcription (RT)-PCR assay was developed for cyclin D1 mRNA suitable for use with RNA extracted from fresh and formalin-fixed, paraffin-embedded tissues. Specimens were amplified in an Applied Biosystems Model 7700 Sequence Detection System in reactions co ntaining primers and probes for cyclin D1 and a control gene, beta (2)-micr oglobulin. Relative expression of the two genes was standardized against a control MCL cell line, M02058. Results: The range of cyclin D1 expression among 20 MCLs was substantially higher than that in other lymphomas and reactive lymph nodes. By choosing a n optimal cutoff point for assessing overexpression, the sensitivity and sp ecificity of the assay for the diagnosis of MCL in lymph node specimens bot h approached 100%: Overexpression was detected in 20 of 20 MCLs, but in non e of 21 non-mantle-cell lymphomas or 10 reactive lymph nodes. Conclusions: Quantitative real-time RT-PCR for cyclin D1 overexpression pro vides a rapid diagnostic test with clinical utility in the diagnosis of MCL . (C) 2001 American Association for Clinical Chemistry.