Cell and Tissue Research

, Volume 333, Issue 3, pp 461–467 | Cite as

Cell-type-specific level of DNA nucleotide excision repair in primary human mammary and ovarian epithelial cell cultures

  • Jean J. Latimer
  • Jennifer M. Johnson
  • Tiffany D. Miles
  • Jason M. Dimsdale
  • Robert P. Edwards
  • Joseph L. Kelley
  • Stephen G. Grant
Regular Article

Abstract

DNA repair, a fundamental function of cellular metabolism, has long been presumed to be constitutive and equivalent in all cells. However, we have previously shown that normal levels of nucleotide excision repair (NER) can vary by 20-fold in a tissue-specific pattern. We have now successfully established primary cultures of normal ovarian tissue from seven women by using a novel culture system originally developed for breast epithelial cells. Epithelial cells in these cultures aggregated to form three-dimensional structures called “attached ovarian epispheres”. The availability of these actively proliferating cell cultures allowed us to measure NER functionally and quantitatively by the unscheduled DNA synthesis (UDS) assay, a clinical test used to diagnose constitutive deficiencies in NER capacity. We determined that ovarian epithelial cells manifested an intermediate level of NER capacity in humans, viz., only 25% of that of foreskin fibroblasts, but still 2.5-fold higher than that of peripheral blood lymphocytes. This level of DNA repair capacity was indistinguishable from that of normal breast epithelial cells, suggesting that it might be characteristic of the epithelial cell type. Similar levels of NER activity were observed in cultures established from a disease-free known carrier of a BRCA1 truncation mutation, consistent with previous normal results shown in breast epithelium and blood lymphocytes. These results establish that at least three “normal” levels of such DNA repair occur in human tissues, and that NER capacity is epigenetically regulated during cell differentiation and development.

Keywords

Primary ovarian epithelial culture Nucleotide excision repair Unscheduled DNA synthesis Xeroderma pigmentosum BRCA1 Human 

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Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Jean J. Latimer
    • 1
    • 3
  • Jennifer M. Johnson
    • 2
  • Tiffany D. Miles
    • 1
  • Jason M. Dimsdale
    • 3
  • Robert P. Edwards
    • 3
  • Joseph L. Kelley
    • 3
  • Stephen G. Grant
    • 1
    • 4
  1. 1.Center for Environmental OncologyUniversity of Pittsburgh Cancer InstitutePittsburghUSA
  2. 2.Program in Biochemistry and Molecular GeneticsUniversity of Pittsburgh School of MedicinePittsburghUSA
  3. 3.Division of Gynecologic Oncology, Department of Obstetrics, Gynecology and Reproductive ScienceUniversity of Pittsburgh School of Medicine and Magee-Womens HospitalPittsburghUSA
  4. 4.Department of Environmental and Occupational Health, Graduate School of Public HealthUniversity of PittsburghPittsburghUSA

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