The role of DNA polymerase eta in translesion synthesis past platinum-DNA adducts in human fibroblasts

Cancer Res. 2004 Sep 15;64(18):6469-75. doi: 10.1158/0008-5472.CAN-04-1328.

Abstract

Cisplatin, a widely used chemotherapeutic agent, has been implicated in the induction of secondary tumors in cancer patients. This drug is presumed to be mutagenic because of error-prone translesion synthesis of cisplatin adducts in DNA. Oxaliplatin is effective in cisplatin-resistant tumors, but its mutagenicity in humans has not been reported. The polymerases involved in bypass of cisplatin and oxaliplatin adducts in vivo are not known. DNA polymerase eta is the most efficient polymerase for bypassing platinum adducts in vitro. We evaluated the role of polymerase eta in translesion synthesis past platinum adducts by determining cytotoxicity and induced mutation frequencies at the hypoxanthine guanine phosphoribosyltransferase (HPRT) locus in diploid human fibroblasts. Normal human fibroblasts (NHF1) were compared with xeroderma pigmentosum variant (XPV) cells (polymerase eta-null) after treatment with cisplatin. In addition, XPV cells complemented for polymerase eta expression were compared with the isogenic cells carrying the empty expression vector. Cytotoxicity and induced mutagenicity experiments were measured in parallel in UVC-irradiated fibroblasts. We found that equitoxic doses of cisplatin induced mutations in fibroblasts lacking polymerase eta at frequencies 2- to 2.5-fold higher than in fibroblasts with either normal or high levels of polymerase eta. These results indicate that polymerase eta is involved in error-free translesion synthesis past some cisplatin adducts. We also found that per lethal event, cisplatin was less mutagenic than UVC. Treatment with a wide range of cytotoxic doses of oxaliplatin did not induce mutations above background levels in cells either expressing or lacking polymerase eta, suggesting that oxaliplatin is nonmutagenic in human fibroblasts.

Publication types

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

MeSH terms

  • Antineoplastic Agents / metabolism
  • Antineoplastic Agents / pharmacology
  • Cisplatin / metabolism
  • Cisplatin / pharmacology*
  • DNA / biosynthesis*
  • DNA / genetics
  • DNA Adducts / metabolism*
  • DNA-Directed DNA Polymerase / biosynthesis
  • DNA-Directed DNA Polymerase / metabolism*
  • Fibroblasts / drug effects
  • Fibroblasts / enzymology*
  • Fibroblasts / physiology
  • Frameshift Mutation
  • Gene Deletion
  • Humans
  • Hypoxanthine Phosphoribosyltransferase / genetics
  • Male
  • Organoplatinum Compounds / metabolism
  • Organoplatinum Compounds / pharmacology*
  • Oxaliplatin
  • Ultraviolet Rays
  • Xeroderma Pigmentosum / genetics
  • Xeroderma Pigmentosum / pathology

Substances

  • Antineoplastic Agents
  • DNA Adducts
  • Organoplatinum Compounds
  • Oxaliplatin
  • DNA
  • Hypoxanthine Phosphoribosyltransferase
  • DNA-Directed DNA Polymerase
  • Rad30 protein
  • Cisplatin