Use of the polymerase chain reaction technique to determine c-myc expression in follicular center cell lymphoma.
We determined, in a semiquantitative fashion, the level of expression of c-myc in 18 follicular center cell lymphomas and five non-neoplastic lymph nodes using reverse transcription and the polymerase chain reaction technique (RT-PCR). With this method, RNA is extracted from lymph node specimens and reverse-transcribed to produce cDNA, which is then amplified using primers specific for c-myc sequences that span introns, thus precluding amplification of genomic DNA. Amplified products are compared with beta2-microglobulin sequences co-amplified in each case as a control for the quality of RNA extracted, RT, and PCR amplification. Using cell lines derived from Burkitt's lymphomas, the RT-PCR method yielded results equivalent to standard Northern blot analysis yet required smaller quantities of tissue. The c-myc transcripts were detected in all lymphoma cases studied (seven high, 11 low expression) and in all non-neoplastic lymph nodes. High or low c-myc expression was based on comparison with non-neoplastic lymph nodes. We conclude that the RT-PCR method is a sensitive, reliable method for measuring gene expression in lymphoma tissues and may be useful for studying the role of c-myc in follicular lymphomas.
Wang
J
J
Laboratory of Pathology, National Cancer Institute, Bethesda, Maryland, USA.
Medeiros
L J
LJ
Longo
D L
DL
Mansoor
A
A
Raffeld
M
M
Duffey
P L
PL
Jaffe
E S
ES
Stetler-Stevenson
M
M
D000328:Adult; D001483:Base Sequence; D015152:Blotting, Northern; D002051:Burkitt Lymphoma / Q000235:genetics / Q000473:pathology; D005260:Female; D016259:Genes, myc; D006801:Humans; D008224:Lymphoma, Follicular / Q000235:genetics* / Q000473:pathology*; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D014407:Tumor Cells, Cultured