Sequence analysis of the 5′-flanking regions of human dihydropyrimidine dehydrogenase gene: Identification of a new polymorphism related with effects of 5-fluorouracil

Takako Hasegawa, Hye-Sook Kim, Masakazu Fukushima, Yusuke Wataya

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Dihydropyrimidine dehydrogenase (DPD), known as a rate-limiting metabolic enzyme in the catabolism of 5-fluorouracil (5-FU), degrades more than about 80% of the administered 5-FU in human liver. Since it was reported that the anticancer effects of 5-FU were observed in cancer patients with lower DPD activities, many attempts have been conducted to anticipate the expected anticancer effects of 5-FU based on expression of intracanceral DPD. It have been reported that 39 different mutations and polymorphisms in the coding regions of DPD genes have been identified; however, there is no report on polymorphisms in the 5-flanking region of DPD genes. We investigated polymorphisms in the 5′-flanking regions (3,058 bp), which are considered to control expression of DPD genes, in genomic DNA extracted from 37 kinds of human cancer cells. As the results, out of 37 cancer cells subjected to analysis, DLD-1 cells had C insertion and 7 strains G deletion, which were hetelozygote. No significant relationship was identified between the DPD activity and the expression levels of DPD mRNA in examined 10 kinds of human cancer cells. However, in DLD-1 cells, which have C-insertion polymorphism in 5′-flanking region of DPD gene, the DPD activity was below detection limit (≤ 0.5 pmol/min/mg protein). Furthermore, 50% of cytosine residue on the CpG site generated by the C insertion was methylated at the 5 position. In this study, we have identified novel polymorphism possibly related the cytotoxicity of 5-FU in the 5′-flanking region of DPD gene. It is suggested that newly identified polymorphism of DPD gene might affect transcription of DPD, thereby providing influence on the clinical outcome of cancer patients treated with 5-FU.

Original languageEnglish
Pages (from-to)233-242
Number of pages10
JournalNucleosides, Nucleotides and Nucleic Acids
Volume24
Issue number4
DOIs
Publication statusPublished - 2005

Fingerprint

Dihydrouracil Dehydrogenase (NADP)
5' Flanking Region
Polymorphism
Fluorouracil
Sequence Analysis
Genes
Cells
Neoplasms
Cytosine
Transcription
Cytotoxicity

Keywords

  • 5-Fluorouracil
  • Dihydropyrimidine Dehydrogenase
  • Polymorphism

ASJC Scopus subject areas

  • Genetics
  • Biochemistry

Cite this

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title = "Sequence analysis of the 5′-flanking regions of human dihydropyrimidine dehydrogenase gene: Identification of a new polymorphism related with effects of 5-fluorouracil",
abstract = "Dihydropyrimidine dehydrogenase (DPD), known as a rate-limiting metabolic enzyme in the catabolism of 5-fluorouracil (5-FU), degrades more than about 80{\%} of the administered 5-FU in human liver. Since it was reported that the anticancer effects of 5-FU were observed in cancer patients with lower DPD activities, many attempts have been conducted to anticipate the expected anticancer effects of 5-FU based on expression of intracanceral DPD. It have been reported that 39 different mutations and polymorphisms in the coding regions of DPD genes have been identified; however, there is no report on polymorphisms in the 5-flanking region of DPD genes. We investigated polymorphisms in the 5′-flanking regions (3,058 bp), which are considered to control expression of DPD genes, in genomic DNA extracted from 37 kinds of human cancer cells. As the results, out of 37 cancer cells subjected to analysis, DLD-1 cells had C insertion and 7 strains G deletion, which were hetelozygote. No significant relationship was identified between the DPD activity and the expression levels of DPD mRNA in examined 10 kinds of human cancer cells. However, in DLD-1 cells, which have C-insertion polymorphism in 5′-flanking region of DPD gene, the DPD activity was below detection limit (≤ 0.5 pmol/min/mg protein). Furthermore, 50{\%} of cytosine residue on the CpG site generated by the C insertion was methylated at the 5 position. In this study, we have identified novel polymorphism possibly related the cytotoxicity of 5-FU in the 5′-flanking region of DPD gene. It is suggested that newly identified polymorphism of DPD gene might affect transcription of DPD, thereby providing influence on the clinical outcome of cancer patients treated with 5-FU.",
keywords = "5-Fluorouracil, Dihydropyrimidine Dehydrogenase, Polymorphism",
author = "Takako Hasegawa and Hye-Sook Kim and Masakazu Fukushima and Yusuke Wataya",
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T1 - Sequence analysis of the 5′-flanking regions of human dihydropyrimidine dehydrogenase gene

T2 - Identification of a new polymorphism related with effects of 5-fluorouracil

AU - Hasegawa, Takako

AU - Kim, Hye-Sook

AU - Fukushima, Masakazu

AU - Wataya, Yusuke

PY - 2005

Y1 - 2005

N2 - Dihydropyrimidine dehydrogenase (DPD), known as a rate-limiting metabolic enzyme in the catabolism of 5-fluorouracil (5-FU), degrades more than about 80% of the administered 5-FU in human liver. Since it was reported that the anticancer effects of 5-FU were observed in cancer patients with lower DPD activities, many attempts have been conducted to anticipate the expected anticancer effects of 5-FU based on expression of intracanceral DPD. It have been reported that 39 different mutations and polymorphisms in the coding regions of DPD genes have been identified; however, there is no report on polymorphisms in the 5-flanking region of DPD genes. We investigated polymorphisms in the 5′-flanking regions (3,058 bp), which are considered to control expression of DPD genes, in genomic DNA extracted from 37 kinds of human cancer cells. As the results, out of 37 cancer cells subjected to analysis, DLD-1 cells had C insertion and 7 strains G deletion, which were hetelozygote. No significant relationship was identified between the DPD activity and the expression levels of DPD mRNA in examined 10 kinds of human cancer cells. However, in DLD-1 cells, which have C-insertion polymorphism in 5′-flanking region of DPD gene, the DPD activity was below detection limit (≤ 0.5 pmol/min/mg protein). Furthermore, 50% of cytosine residue on the CpG site generated by the C insertion was methylated at the 5 position. In this study, we have identified novel polymorphism possibly related the cytotoxicity of 5-FU in the 5′-flanking region of DPD gene. It is suggested that newly identified polymorphism of DPD gene might affect transcription of DPD, thereby providing influence on the clinical outcome of cancer patients treated with 5-FU.

AB - Dihydropyrimidine dehydrogenase (DPD), known as a rate-limiting metabolic enzyme in the catabolism of 5-fluorouracil (5-FU), degrades more than about 80% of the administered 5-FU in human liver. Since it was reported that the anticancer effects of 5-FU were observed in cancer patients with lower DPD activities, many attempts have been conducted to anticipate the expected anticancer effects of 5-FU based on expression of intracanceral DPD. It have been reported that 39 different mutations and polymorphisms in the coding regions of DPD genes have been identified; however, there is no report on polymorphisms in the 5-flanking region of DPD genes. We investigated polymorphisms in the 5′-flanking regions (3,058 bp), which are considered to control expression of DPD genes, in genomic DNA extracted from 37 kinds of human cancer cells. As the results, out of 37 cancer cells subjected to analysis, DLD-1 cells had C insertion and 7 strains G deletion, which were hetelozygote. No significant relationship was identified between the DPD activity and the expression levels of DPD mRNA in examined 10 kinds of human cancer cells. However, in DLD-1 cells, which have C-insertion polymorphism in 5′-flanking region of DPD gene, the DPD activity was below detection limit (≤ 0.5 pmol/min/mg protein). Furthermore, 50% of cytosine residue on the CpG site generated by the C insertion was methylated at the 5 position. In this study, we have identified novel polymorphism possibly related the cytotoxicity of 5-FU in the 5′-flanking region of DPD gene. It is suggested that newly identified polymorphism of DPD gene might affect transcription of DPD, thereby providing influence on the clinical outcome of cancer patients treated with 5-FU.

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