Fujiwara Hisataka, Terashima Masanori, Irinoda Takashi, Takagane Akinori, Abe Kaoru, Kashiwaba Masahiro, Oyama Kennichi, Takahashi Masanori, Maesawa Chihaya, Saito Kazuyoshi, Takechi Teiji, Fukushima Masakazu
Department of Surgery, Iwate Medical University, Morioka 020-8505, Japan.
Jpn J Cancer Res. 2002 Dec;93(12):1342-50. doi: 10.1111/j.1349-7006.2002.tb01243.x.
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.
我们使用实时逆转录聚合酶链反应（RT-PCR）来检测胃癌组织中胸苷酸合成酶（TS）和二氢嘧啶脱氢酶（DPD）的mRNA表达，目的是建立一个用于检测超低体积样本中TS和DPD的系统。本研究以5种人胃癌细胞系的裸鼠异种移植瘤和85份临床样本作为标本。根据小鼠体内相对肿瘤增殖率以及使用临床样本无血清培养物进行ATP检测的结果来确定对5-氟尿嘧啶（5-FU）的敏感性。使用ABI PRISM 7700系统通过实时RT-PCR测量肿瘤组织中的mRNA表达。TS和DPD的mRNA表达值根据甘油醛-3-磷酸脱氢酶mRNA表达水平进行校正。异种移植瘤显示TS和DPD的mRNA表达与酶活性之间存在相关性（TS：rs = 0.700，DPD：rs = 0.900），并且mRNA水平与对5-FU的敏感性之间存在负相关（TS：rs = -0.900，DPD：rs = -0.800）。临床样本显示5-FU敏感性与mRNA表达之间存在负相关（TS：rs = -0.518，DPD：rs = -0.564）。仅在TS mRNA表达和DPD mRNA表达均较低的病例中观察到对5-FU的敏感性。实时RT-PCR可以对胃癌中TS和DPD的mRNA表达进行高度灵敏的评估，并且对于预测5-FU敏感性很有用。
