Real-time quantitative reverse transcription-PCR for cyclin D1 mRNA in blood, marrow, and tissue specimens for diagnosis of mantle cell lymphoma.
BACKGROUND
Overexpression of cyclin D1 mRNA, found in mantle cell lymphoma (MCL), is a critical diagnostic marker. We investigated the use of real-time reverse transcription-PCR (RT-PCR) for cyclin D1.


METHODS
We studied 97 fresh specimens (50 blood, 30 bone marrow, 15 lymph node, and 2 other samples) from patients diagnosed with a variety of lymphoproliferative diseases, including 25 cases of MCL. We used real-time quantitative RT-PCR to evaluate cyclin D1 mRNA expression. Because blood and marrow specimens may contain only a minority of potentially malignant cells (as opposed to most lymph nodes) and to increase sensitivity, we normalized the cyclin D1 mRNA concentrations to mRNA of a B-cell-specific marker, CD19, as well as to previously characterized beta(2)-microglobulin mRNA.


RESULTS
In 16 of 21 cases of MCL with overt disease, the ratio of cyclin D1 mRNA to beta(2)-microglobulin mRNA was increased, but all 21 cases showed increased ratios of cyclin D1 mRNA to CD19 mRNA. Cyclin D1 mRNA was low or undetectable in various lymphoproliferative diseases, including cases of ambiguous immunophenotype. The mRNA ratios were stable over 3-7 days of sample storage.


CONCLUSIONS
Quantitative RT-PCR for cyclin D1 mRNA normalized to CD19 mRNA can be used in the diagnosis of MCL in blood, marrow, and tissue.
Howe
John Greg
JG
Department of Laboratory Medicine, Yale University School of Medicine, 333 Cedar St., New Haven, CT 06520, USA.
Crouch
Jill
J
Cooper
Dennis
D
Smith
Brian R
BR
D018941:Antigens, CD19 / Q000096:biosynthesis / Q000097:blood / Q000235:genetics; D014408:Biomarkers, Tumor / Q000032:analysis* / Q000097:blood; D001853:Bone Marrow / Q000378:metabolism*; D019938:Cyclin D1 / Q000096:biosynthesis* / Q000097:blood / Q000235:genetics; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D006801:Humans; D016130:Immunophenotyping; D008198:Lymph Nodes / Q000378:metabolism; D020522:Lymphoma, Mantle-Cell / Q000175:diagnosis / Q000378:metabolism* / Q000473:pathology; D012333:RNA, Messenger / Q000096:biosynthesis* / Q000097:blood / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D013997:Time Factors; D001613:beta 2-Microglobulin / Q000096:biosynthesis / Q000097:blood / Q000235:genetics
D018941:Antigens, CD19; D014408:Biomarkers, Tumor; D012333:RNA, Messenger; D001613:beta 2-Microglobulin; D019938:Cyclin D1