Association between the level of ERCC-1 expression and the repair of cisplatin-induced DNA damage in human ovarian cancer cells

Anticancer Res. 2000 Mar-Apr;20(2A):645-52.

Abstract

Nucleotide excision repair (NER) is responsible for the repair of platinum-DNA lesions. ERCC-1 is a critical gene within the NER pathway, and cells without a functional ERCC-1 do not repair cisplatin-caused DNA damage. The present study was therefore designed to evaluate the relationship between the expression of ERCC-1 and the repair of cisplatin-induced DNA adducts in human ovarian cancer cells in vitro. One hour exposure of MCAS cells to cisplatin yielded an approximately two-fold increment in the levels of ERCC-1 mRNA and ERCC-1 protein, as determined, respectively, by Northern and Western blottings. In addition, nuclear run-on assay showed that ERCC-1 gene transcription rate was increased to about the same extent as steady-state ERCC-1 mRNA and protein, in response to cisplatin treatment. However, the levels of ERCC-1 mRNA, ERCC-1 protein, and ERCC-1 transcript in MCAS cells are two-fold lower than those in A2780/CP70 cells, as previously reported. Furthermore, the repair of cisplatin-DNA adducts in MCAS cells, as measured by atomic absorption spectrometry, is also nearly two-fold less than that in A2780/CP70 cells, indicating a strong association between the level of ERCC-1 expression and the activity of excision repair in these two human ovarian tumor cell lines. These results suggest that ERCC-1 may be a useful marker to monitor the repair of platinum-DNA damage in tumor cells, and further highlight that potential pharmacological approaches which specifically inhibit ERCC-1 expression may increase cellular sensitivity to cisplatin.

Publication types

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

MeSH terms

  • Cell Nucleus / metabolism
  • Cisplatin / analysis*
  • Cisplatin / toxicity*
  • DNA Adducts / analysis*
  • DNA Damage*
  • DNA Repair*
  • DNA-Binding Proteins*
  • Endonucleases*
  • Female
  • Gene Expression Regulation, Neoplastic* / drug effects
  • Humans
  • Kinetics
  • Ovarian Neoplasms
  • Proteins / genetics*
  • RNA, Messenger / genetics
  • Transcription, Genetic
  • Tumor Cells, Cultured

Substances

  • DNA Adducts
  • DNA-Binding Proteins
  • Proteins
  • RNA, Messenger
  • cisplatin-DNA adduct
  • ERCC1 protein, human
  • Endonucleases
  • Cisplatin