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Carcinogenesis

Increased Resistance to N,N′,N″-Triethylenethiophosphoramide (Thiotepa) in Cells Expressing the Escherichia coli Formamidopyrimidine-DNA Glycosylase

R. D. Gill, C. Cussac, R. L. Souhami and F. Laval
R. D. Gill
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C. Cussac
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R. L. Souhami
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F. Laval
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DOI:  Published August 1996
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Abstract

Thiotepa (N,N',N"-triethylenethiophosphoramide) is an alkylating agent used in cancer chemotherapy. A reaction pathway by which thiotepa alkylates purified DNA involves hydrolysis to aziridine, which forms N7-aminoethyl guanine and aminoethyl adenine. These lesions are repaired in Escherichia coli by the formamidopyrimidine-DNA glycosylase (Fpg) protein. To determine whether such lesions are formed by thiotepa in mammalian cells, we have overexpressed the E. coli Fpg protein in Chinese hamster ovary cells. The transfected cells were more resistant to the lethal and mutagenic effects of thiotepa than the parental cells. The number of replication-blocking lesions formed by thiotepa, measured by quantitative PCR analysis, was lower in the transfected cells. These results show that expression of the Fpg protein increases the cell resistance to thiotepa and suggest that this compound produces ring-opened guanines, which are involved in its cytotoxic action.

Footnotes

  • ↵1 This work was supported by grants from the Institut National de la Santé et de la Recherche Médicale; ARC (Villejuif); the Special Trustees of Middlesex Hospital, London; and the Cancer Research Campaign (CRC), United Kingdom.

  • ↵2 To whom requests for reprints should be addressed. Phone: 33-1-49-59-18-53; Fax: 33-1-45-59-48-24.

  • Received March 29, 1996.
  • Accepted June 18, 1996.
  • ©1996 American Association for Cancer Research.
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August 1996
Volume 56, Issue 16
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Increased Resistance to N,N′,N″-Triethylenethiophosphoramide (Thiotepa) in Cells Expressing the Escherichia coli Formamidopyrimidine-DNA Glycosylase
R. D. Gill, C. Cussac, R. L. Souhami and F. Laval
Cancer Res August 15 1996 (56) (16) 3721-3724;

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Increased Resistance to N,N′,N″-Triethylenethiophosphoramide (Thiotepa) in Cells Expressing the Escherichia coli Formamidopyrimidine-DNA Glycosylase
R. D. Gill, C. Cussac, R. L. Souhami and F. Laval
Cancer Res August 15 1996 (56) (16) 3721-3724;
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