Basic Studies on Prediction of Adverse Reactins to Pyrimidine Anti-Cancer Drugs-Analyses on Pyrimidine Metaboism Disonders-

1. 二氢嘧啶酶mrna的分析二氢嘧啶酶基因组DNA有10个外显子，分析过程复杂。我们可以用ISOGEN从外周血中分离mRNA。亚克隆后，我们对cDNA进行测序。我们在二氢嘧啶尿患者中发现了报道的突变，表明这种方法在临床上是有用的。我们分析了两例氟尿嘧啶严重中毒患者的二氢嘧啶脱氢酶基因。并发现了新的突变。患者1有1003G→T)突变和1156G→T突变。我们诊断患者有DPD缺陷，并推测这些突变与严重的氟尿嘧啶毒性有关。患者2有1626A→G突变，但我们不能明确这种突变与严重的氟尿嘧啶毒性之间的关系。

1. Analysis on dihydropyrimidinase mRNAAnalysis on the dihydropyrimidinase genomic DNA needs complicated procedures because it has 10 exons. We could separated mRNA from peripheral blood, using ISOGEN.After subcloned, we sequenced cDNA.We identified the reported mutations In the patient with dihydropyrimidinuria, suggesting that this method is clinically useful.2. Analysis on dihydropyrimidine dehydrogenase in two patient with a severe fluorouracil toxicityWe analysed the dihydropyrimidine dehydrogenase gene in two patient with a severe fluorouracil toxicity. and identified novel mutations. The patient 1 had a 1003G→T) mutation and a 1156G→Tmutation. We diagnosed the patient as having DPD deflciency, and supeculated that these mutations were associated with a severe fluorouracil toxicity.The patient 2 had a 1626A→G mutation, but we could not clarify the relationship between this mutation and a severe fluorouracil toxicity.