RRC ID |
61969
|
著者 |
Fujiwara H, Terashima M, Irinoda T, Takagane A, Abe K, Kashiwaba M, Oyama K, Takahashi M, Maesawa C, Saito K, Takechi T, Fukushima M.
|
タイトル |
Quantitative measurement of thymidylate synthase and dihydropyrimidine dehydrogenase mRNA level in gastric cancer by real-time RT-PCR.
|
ジャーナル |
Jpn J Cancer Res
|
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.
|
巻・号 |
93(12)
|
ページ |
1342-50
|
公開日 |
2002-12-1
|
DOI |
10.1111/j.1349-7006.2002.tb01243.x
|
PMID |
12495474
|
PMC |
PMC5926932
|
MeSH |
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
Stomach Neoplasms / enzymology*
Thymidylate Synthase / genetics*
Transplantation, Heterologous
Tumor Cells, Cultured
|
リソース情報 |
ヒト・動物細胞 |
MKN1(RCB1003)
MKN74(RCB1002)
GCIY(RCB0555) |