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

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.

• Authors: Hisataka Fujiwara, Masanori Terashima, Takashi Irinoda, Akinori Takagane, Kaoru Abe, Masahiro Kashiwaba, Kennichi Oyama, Masanori Takahashi, Chihaya Maesawa, Kazuyoshi Saito, Teiji Takechi, Masakazu Fukushima
• Journal: Japanese journal of cancer research : Gann (Jpn J Cancer Res) Vol. 93 Issue 12 Pg. 1342-50 (Dec 2002) ISSN: 0910-5050 [Print] Japan
• PMID: 12495474 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• RNA, Messenger
• Oxidoreductases
• Dihydrouracil Dehydrogenase (NADP)
• Thymidylate Synthase
• Fluorouracil

Topics

• Animals
• Dihydrouracil Dehydrogenase (NADP)
• Female
• Fluorouracil (pharmacology)
• Humans
• Male
• Mice
• Mice, Inbred BALB C
• Neoplasm Transplantation
• Oxidoreductases (genetics)
• RNA, Messenger (analysis)
• Reverse Transcriptase Polymerase Chain Reaction (methods)
• Stomach Neoplasms (drug therapy, enzymology)
• Thymidylate Synthase (genetics)
• Transplantation, Heterologous
• Tumor Cells, Cultured