BACKGROUND: Measurement of intratumoral expression of dihydropyrimidine dehydrogenase (DPD) and thymidylate synthase (TS) may have some value in predicting the response to fluoropyrimidine-containing therapy. PATIENTS AND METHODS: We attempted to validate this association in a series of Russian metastatic colorectal cancer cases. While replicating already published protocols, we unexpectedly found that the use of commonly utilized gene referees, glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and beta-actin, may lead to artifacts due to pseudogene-driven amplification from the genomic DNA template. We have developed a real-time PCR protocol which amplifies short PCR fragments, thus allowing efficient analysis of archival formalin-fixed paraffin-embedded tumor samples, and relies on succinate dehydrogenase (SDHA) as a gene referee, therefore avoiding amplification from genomic DNA. RESULTS: Low content of DPD transcripts was observed in 13/20 (65%) patients with disease control (tumor response or disease stabilization) as compared to only 3/9 (33%) subjects with progressive disease (p = 0.11). Despite the low number of patients, this association reached the level of statistical significance when similar analysis was done for TS expression (11/20 (55%) vs. 1/9 (11%); p = 0.03). CONCLUSIONS: Our data confirm that low DPD and TS expressors have higher chances of success of fluoropyrimidine-containing regimens.
BACKGROUND: Measurement of intratumoral expression of dihydropyrimidine dehydrogenase (DPD) and thymidylate synthase (TS) may have some value in predicting the response to fluoropyrimidine-containing therapy. PATIENTS AND METHODS: We attempted to validate this association in a series of Russian metastatic colorectal cancer cases. While replicating already published protocols, we unexpectedly found that the use of commonly utilized gene referees, glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and beta-actin, may lead to artifacts due to pseudogene-driven amplification from the genomic DNA template. We have developed a real-time PCR protocol which amplifies short PCR fragments, thus allowing efficient analysis of archival formalin-fixed paraffin-embedded tumor samples, and relies on succinate dehydrogenase (SDHA) as a gene referee, therefore avoiding amplification from genomic DNA. RESULTS: Low content of DPD transcripts was observed in 13/20 (65%) patients with disease control (tumor response or disease stabilization) as compared to only 3/9 (33%) subjects with progressive disease (p = 0.11). Despite the low number of patients, this association reached the level of statistical significance when similar analysis was done for TS expression (11/20 (55%) vs. 1/9 (11%); p = 0.03). CONCLUSIONS: Our data confirm that low DPD and TS expressors have higher chances of success of fluoropyrimidine-containing regimens.
Authors: Irina O Vassilieva; Victor N Tomilin; Irina I Marakhova; Alla N Shatrova; Yuri A Negulyaev; Svetlana B Semenova Journal: J Membr Biol Date: 2012-11-01 Impact factor: 1.843
Authors: Evgeny N Suspitsin; Aniruddh Kashyap; Kseniya V Shelekhova; Anna P Sokolenko; Ekatherina Sh Kuligina; Aglaya G Iyevleva; Alexandr V Kornilov; Volker Ehemann; Grigoriy A Yanus; Svetlana N Aleksakhina; Elena V Preobrazhenskaya; Olga A Zaitseva; Olga S Yatsuk; Valeriy F Klimashevsky; Alexandr V Togo; Evgeny N Imyanitov Journal: Med Oncol Date: 2013-06-26 Impact factor: 3.064
Authors: Vladimir M Moiseyenko; Nikita M Volkov; Evgeny N Suspistin; Grigoriy A Yanus; Aglaya G Iyevleva; Ekatherina Sh Kuligina; Alexandr V Togo; Alexandr V Kornilov; Alexandr O Ivantsov; Evgeny N Imyanitov Journal: Med Oncol Date: 2013-03-27 Impact factor: 3.064