Use of the Polymerase Chain Reaction Technique to Determine c-myc Expression in Follicular Center Cell Lymphoma

Abstract

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.

No correlation of c- myc overexpression and p53 mutations in liposarcomas

“…c-myc expression level was analysed by differential RT-PCR. Reliability and validity of the RT-PCR method were demonstrated earlier for c-myc [41,50], for the mdr1 gene [34], and for mdm2 [39]. We found elevated levels of c-myc expression in 6 of 31 liposarcomas (19.3%) investigated.…”

“…sense: 5′TCGCAAGACTCCAGCGCCTTCTC-3′ and antisense: 5′-TGACACTGTCCAACTTGACCCTCTT-3′, which yielded a 363-bp product [50]. Oligonucleotides used for the β-2MG-specific sequence as internal control were sense: 5′-TTAGCTGT-GCTCGCGCTACTCTCT-3′ and antisense: 5′-GTCGGATGGAT-GAAACCCAGACAC-3′, which yielded a 144-bp product.…”