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Detection of cyclin D1 (bcl-1, PRAD1) overexpression by a simple
competitive reverse transcription-polymerase chain reaction assay in
t(11;14)(q13;q32)-bearing B-cell malignancies and/or mantle cell lymphoma
K Uchimaru, T Taniguchi, M Yoshikawa, S Asano, A Arnold, T Fujita and T Motokura
Fourth Department of Internal Medicine, School of Medicine, University of
Tokyo, Japan.
In mantle cell lymphoma, the t(11;14)(q13;q32) and its molecular
counterpart, bcl-1 rearrangement, are consistent features and lead to
cyclin D1 (bcl-1, PRAD1) proto-oncogene overexpression. In order to detect
cyclin D1 overexpression, we developed a simple assay involving a reverse
transcription followed by competitive polymerase chain reaction (PCR). A
single upstream primer was derived from a homologous region between cyclin
D1 and the other D-type cyclins, cyclins D2 and D3, while three downstream
primers were specific to their respective D- type cyclins. Because the
upstream primer was shared in PCR amplification of the three sequences,
each PCR product served as a competitor and the quantification of the
target was made by comparison of the intensity of the three products. With
this assay we analyzed 45 hematopoietic cell lines and 40 clinical
specimens. Cyclin D1 was rarely expressed in lymphoid cell lines except in
t(11;14)(q13;q32)- bearing B-cell malignancies and/or mantle cell lymphoma,
which expressed cyclin D1 predominantly. In myeloid cell lines, the levels
of cyclin D1 expression varied and never exceeded the sum of cyclin D2 and
D3 levels. Cyclin D3 was ubiquitously expressed while cyclins D1 and D2
were differentially used. The observations suggest that human cyclin D3 may
play a fundamental role in hematopoiesis and that cyclins D1 and D2 may
have different lineage- or differentiation-dependent functions. With this
assay, small aliquots of clinical specimens such as 100 microL peripheral
blood were enough to detect cyclin D1 overexpression without a
well-controlled standard. The technique was validated as highly comparable
with Northern analysis. This rapid and reliable detection of cyclin D1
overexpression may have practical clinical utility in the analysis and
management of B-cell malignancies.
Volume 89,
Issue 3,
pp. 965-974,
02/01/1997
Copyright © 1997 by The American Society of Hematology
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