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Related Experiment Videos

Amsacrine-induced mutations in AS52 cells

L R Ferguson1, P M Turner, D W Hart

  • 1Cancer Research Laboratory, University of Auckland Medical School, New Zealand.

Environmental and Molecular Mutagenesis
|August 26, 1998
PubMed
Summary
This summary is machine-generated.

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Amsacrine treatment of AS52 cells induced mutations, primarily genomic deletions and rearrangements in the gpt gene. These alterations were confirmed through PCR, Southern, and DNA sequencing, highlighting their role in drug resistance.

Area of Science:

  • Molecular Biology
  • Genetics
  • Toxicology

Background:

  • Amsacrine is a topoisomerase II inhibitor that intercalates into DNA.
  • AS52 cells contain a single copy of the bacterial gpt gene integrated into the Chinese hamster ovary genome.

Purpose of the Study:

  • To analyze the molecular mechanisms of 6-thioguanine resistance in AS52 cells after amsacrine treatment.
  • To characterize mutations in the gpt gene induced by amsacrine.

Main Methods:

  • AS52 cells were treated with amsacrine (0.1-0.5 microM).
  • 6-thioguanine resistant colonies were isolated and analyzed using PCR, Southern blotting, Northern blotting, and DNA sequencing.

Main Results:

  • Amsacrine treatment resulted in significant cytotoxicity and mutagenicity.

Related Experiment Videos

  • Genomic deletions and rearrangements were the most common mutations, observed in 12/21 mutants analyzed by PCR and Southern blotting.
  • Mutations affected gpt gene expression, with 4 out of 9 sequenced mutants having alterations in the gpt structural gene.

    • Conclusions:

    • Genomic deletions and rearrangements are the predominant mutation types induced by amsacrine in AS52 cells.
    • These alterations play a significant role in conferring 6-thioguanine resistance.