Quantitative RNA slot-blot analysis of CCND1/cyclin D1 expression in suspected mantle cell lymphoma.
Abnormal CCND1 expression is found in the majority of mantle cell lymphomas (MCL) and in a minority of other mature B cell malignancies. Its evaluation can therefore aid diagnostic classification, in conjunction with clinical, morphological, immunophenotypic and cytogenetic analysis. We describe a rapid slot-blot hybridization technique allowing quantitative assessment of CCND1 expression relative to beta-actin, with a sensitivity cut-off of approximately 10%. This allowed clear separation (P < 0.01) of CCND1 MCL (0.89 +/- 0.4; range 0.23-1.81; n = 25) from control samples (0.02 +/- 0.04; range 0-0.09; n = 22) on limited quantities of RNA (1-3.5 microg). Of nine samples in which a potential diagnosis of MCL lymphoma was based on morphological analysis of paraffin-embedded material, without adequate immunophenotype analysis, all were CCND1 negative and subsequent immunophenotype demonstrated features compatible with chronic lymphocytic leukemia (CLL)/small lymphocytic lymphoma (SLL) (CD5+, CD23+, FMC7-) in all cases tested. This study demonstrates the feasibility of slot-blot CCND1 quantification and the importance of the availability of cryopreserved material.
Mauvieux
L
L
Department of Biological Hematology, CHU Necker Enfants-Malades and Université Paris V, France.
Canioni
D
D
Hermine
O
O
Valensi
F
F
Radford-Weiss
I
I
Azagury
M
M
Magen
M
M
Flandrin
G
G
Brousse
N
N
Varet
B
B
Macintyre
E A
EA
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001345:Autoradiography; D015152:Blotting, Northern / Q000379:methods; D001853:Bone Marrow / Q000473:pathology; D015925:Cryopreservation; D019938:Cyclin D1 / Q000032:analysis / Q000096:biosynthesis*; D003937:Diagnosis, Differential; D005260:Female; D006801:Humans; D016130:Immunophenotyping; D008198:Lymph Nodes / Q000473:pathology; D008214:Lymphocytes / Q000276:immunology / Q000473:pathology; D008228:Lymphoma, Non-Hodgkin / Q000097:blood / Q000276:immunology / Q000473:pathology*; D008297:Male; D008875:Middle Aged; D016066:Pleural Effusion, Malignant / Q000276:immunology / Q000473:pathology; D012680:Sensitivity and Specificity
D019938:Cyclin D1