Gene-specific formation and repair of cisplatin intrastrand adducts and interstrand cross-links in Chinese hamster ovary cells

J Biol Chem. 1991 Apr 15;266(11):7101-7.

Abstract

We have used three methods to study the formation and repair of intrastrand adducts and interstrand cross-links in the DNA of Chinese hamster ovary cells induced by the anticancer drug cis-diamminedichloroplatinum II (cisplatin). Using atomic absorption spectroscopy, we found that 21% of the total genomic cisplatin adducts were removed at 8 h and 42% at 24 h. We used ABC excinuclease digestion, coupled with out previously reported methodology to quantify DNA in specific genomic regions. These adducts were removed faster in the transcribed dihydrofolate reductase and c-myc genes compared to a noncoding fragment, a region containing the little or nontranscribed c-fos oncogene, and to the overall genome. Interstrand cross-links in specific sequences were quantified by Southern hybridization of denatured-renatured DNA separated on a neutral gel. We found that cross-links were removed more efficiently from the gene regions than intrastrand adducts and, at high levels of cross-linking, removal was similar from transcribed and from nontranscribed regions.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Cell Line
  • Cisplatin / pharmacology*
  • Cricetinae
  • Cricetulus
  • Cross-Linking Reagents
  • DNA / drug effects*
  • DNA / isolation & purification
  • DNA Damage*
  • DNA Repair*
  • Female
  • Genes*
  • Ovary
  • Proto-Oncogenes
  • Tetrahydrofolate Dehydrogenase / genetics

Substances

  • Cross-Linking Reagents
  • DNA
  • Tetrahydrofolate Dehydrogenase
  • Cisplatin