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Quantitative measurement of thymidylate synthase and dihydropyrimidine dehydrogenase mRNA level in gastric cancer by real-time RT-PCR.

Authors
  • Fujiwara, Hisataka1
  • Terashima, Masanori
  • Irinoda, Takashi
  • Takagane, Akinori
  • Abe, Kaoru
  • Kashiwaba, Masahiro
  • Oyama, Kennichi
  • Takahashi, Masanori
  • Maesawa, Chihaya
  • Saito, Kazuyoshi
  • Takechi, Teiji
  • Fukushima, Masakazu
  • 1 Department of Surgery, Iwate Medical University, Morioka 020-8505, Japan.
Type
Published Article
Journal
Japanese journal of cancer research : Gann
Publication Date
December 2002
Volume
93
Issue
12
Pages
1342–1350
Identifiers
PMID: 12495474
Source
Medline
License
Unknown

Abstract

We used real-time reverse-transcription polymerase chain reaction (RT-PCR) to assay expression of the mRNA of thymidylate synthase (TS) and dihydropyrimidine dehydrogenase (DPD) in gastric cancer tissue with the objective of establishing a system to measure TS and DPD in ultra-low-volume samples. Nude mouse xenografts of 5 human gastric cancer cell lines and 85 clinical samples were used as the specimens in this study. Sensitivity to 5-fluorouracil (5-FU) was determined on the basis of the relative tumor proliferation rate in mice and the results of ATP assay using serum-free cultures of the clinical samples. mRNA expression was measured in tumor tissue by real-time RT-PCR using the ABI PRISM 7700 system. The values for expression of the mRNA for TS and DPD were corrected according to the level of glyceraldehyde-3-phosphate dehydrogenase mRNA expression. The xenografts yielded correlations between TS and DPD mRNA expression and the activity of the enzymes (TS: rs=0.700, DPD: rs=0.900), and an inverse correlation was noted between the mRNA levels and sensitivity to 5-FU (TS: rs=-0.900, DPD: rs=-0.800). The clinical samples showed an inverse correlation between 5-FU sensitivity and mRNA expression (TS: rs=-0.518, DPD: rs=-0.564). Sensitivity to 5-FU was noted only in cases in which TS mRNA expression and DPD mRNA expression were both low. Real-time RT-PCR can provide a highly sensitive assessment of TS and DPD mRNA expression in gastric cancer, and it was useful for predicting 5-FU sensitivity.

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