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Additive Effects of Bleomycin and Neocarzinostatin on Degradation of DNA, Inhibition of DNA Polymerase β, and Cell Growth

Werner E. G. Müller, Armin Maidhof, Josef Arendes, Werner Geurtsen, Rudolf K. Zahn and Ralf Schmidseder
Werner E. G. Müller
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Armin Maidhof
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Josef Arendes
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Werner Geurtsen
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Rudolf K. Zahn
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Ralf Schmidseder
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DOI:  Published September 1979
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Abstract

The interactions of the two antitumor protein antibiotics, neocarzinostatin (NCS) and bleomycin (BLM), were studied on subcellular and cellular levels.

BLM and NCS were found to remove thymine from double-stranded DNA. Combination experiments using BLM and NCS together in an assay with isolated DNA revealed an additive effect in splitting. Under limiting concentration conditions, BLM and NCS induce alkali-labile sites in DNA without a subsequent cleavage of the chain. After transfer of BLM- or NCS-treated DNA into an alkaline solution, strand scissions occur. Combination of BLM and NCS results in an additive DNA-cleaving effect, which indicates that the splitting reactions initiated by BLM or NCS are not influenced if the two antibiotics are applied in combination.

The DNA polymerase β is inhibited by BLM (at higher concentrations) and by NCS in a competitive way with respect to DNA. The inhibition constant of BLM and NCS in a combination experiment was found to be the result of the sum of the inhibition constants of BLM and NCS.

Using L5178Y mouse lymphoma cells, it was found that cells incubated with both BLM and NCS show “unbalanced growth.” The dose-response curves from BLM and NCS have identical slopes; they are characteristic for compounds which selectively inhibit DNA synthesis. By use of isobolograms, it could also be clearly shown that BLM and NCS interact additively.

Footnotes

  • ↵1 Supported by grants from the Akademie der Wissenschaften und der Literatur, Mainz, Germany, and from the Deutsche Forschungsgemeinschaft. The generous gifts of bleomycin and herring sperm DNA from Fa. H. Mack, Illertissen, Germany and of Neocarzinostatin from Dr. W. T. Bradner (Bristol Laboratories, East Syracuse, N. Y.) are appreciated.

  • ↵2 To whom requests for reprints should be addressed.

  • Received February 5, 1979.
  • Accepted May 23, 1979.
  • ©1979 American Association for Cancer Research.
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September 1979
Volume 39, Issue 9
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Additive Effects of Bleomycin and Neocarzinostatin on Degradation of DNA, Inhibition of DNA Polymerase β, and Cell Growth
Werner E. G. Müller, Armin Maidhof, Josef Arendes, Werner Geurtsen, Rudolf K. Zahn and Ralf Schmidseder
Cancer Res September 1 1979 (39) (9) 3768-3773;

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Additive Effects of Bleomycin and Neocarzinostatin on Degradation of DNA, Inhibition of DNA Polymerase β, and Cell Growth
Werner E. G. Müller, Armin Maidhof, Josef Arendes, Werner Geurtsen, Rudolf K. Zahn and Ralf Schmidseder
Cancer Res September 1 1979 (39) (9) 3768-3773;
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