title: Establishment and characterization of a breast cell strain containing a BRCA1 185delAG mutation.
abstract: OBJECTIVE
  The aim of this study was to examine whether cells containing the heterozygous form of a BRCA1 185delAG mutation would exhibit abnormal growth or an altered response to DNA damage.
  
  
  METHODS
  A primary culture of human mammary epithelial cells (90P) was obtained from the nontumor breast tissue of a 35-year-old patient who had undergone a mastectomy for removal of a breast tumor. These cells were immortalized (90PE6E7) following retroviral infection with HPV-16 viral E6/E7. genes. Both the 90P cell strain and the cell line were characterized for their ability to grow in culture, form colonies in soft agar, and produce tumors in athymic nude mice compared to normal breast epithelial cells containing wild-type BRCA1. 90P cells were also analyzed for cellular response to gamma radiation and H(2)O(2).
  
  
  RESULTS
  These cells were confirmed to contain a frameshift mutation, 185delAG, of the BRCA1 gene. Despite being heterozygous for wild-type BRCA1, the 220-kDa full-size BRCA1 protein was abundantly expressed. 90P and 90PE6E7 cells grew at a similar rate and were anchorage dependent. 90PE6E7 also failed to form tumors in athymic nude mice. Finally, 90P cells exhibited a survival response similar to that of normal mammary epithelial cells to radiation damage and exposure to oxidative stress.
  
  
  CONCLUSIONS
  To our knowledge the 90P cells and the 90PE6E7 cells are the first characterized, non-tumor-derived breast epithelial cells that are heterozygous for the BRCA1 germline mutation 185delAG. Our conclusion is that these BRCA1 mutant cells appear to have growth and stress response characteristics similar to those of normal human breast cells, which is consistent with the hypothesis that loss of heterozygosity must occur to impair putative BRCA1 function.
authors.affiliation: Laboratory of Molecular Carcinogenesis, National Institute of Environmental Health Sciences, Research Triangle Park, North Carolina 27709, USA. Annab@NIEHS.NIH.GOV
authors.forename: L A
authors.initials: LA
authors.lastname: Annab
authors.forename: L
authors.initials: L
authors.lastname: Terry
authors.forename: P L
authors.initials: PL
authors.lastname: Cable
authors.forename: J
authors.initials: J
authors.lastname: Brady
authors.forename: M R
authors.initials: MR
authors.lastname: Stampfer
authors.forename: J C
authors.initials: JC
authors.lastname: Barrett
authors.forename: C A
authors.initials: CA
authors.lastname: Afshari
mesh_terms: D000328:Adult; D000818:Animals; D001943:Breast Neoplasms / Q000235:genetics* / Q000473:pathology; D002471:Cell Transformation, Neoplastic; D003470:Culture Media; D004249:DNA Damage; D005260:Female; D016368:Frameshift Mutation; D019398:Genes, BRCA1 / Q000235:genetics*; D018095:Germ-Line Mutation; D006801:Humans; D019656:Loss of Heterozygosity; D051379:Mice; D008819:Mice, Nude; D014407:Tumor Cells, Cultured
chemical_list: D003470:Culture Media