Localization of BRCA1 protein in breast cancer tissue and cell lines with mutations

Natalie Tulchin, Leonard Ornstein, Steven Dikman, James Strauchen, Shabnam Jaffer, Chandandeep Nagi, Ira Bleiweiss, Ruth Kornreich, Lisa Edelmann, Karen Brown, Carol Bodian, Venugopalan D. Nair, Monique Chambon, Nicholas T. Woods, Alvaro N.A. Monteiro

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12 Scopus citations

Abstract

Background: The breast and ovarian cancer susceptibility gene (BRCA1) encodes a tumor suppressor. The BRCA1 protein is found primarily in cell nuclei and plays an important role in the DNA damage response and transcriptional regulation. Deficiencies in DNA repair capabilities have been associated with higher histopathological grade and worse prognosis in breast cancer.Methods: In order to investigate the subcellular distribution of BRCA1 in tumor tissue we randomly selected 22 breast carcinomas and tested BRCA1 protein localization in frozen and contiguous formalin-fixed, paraffin embedded (FFPE) tissue, using pressure cooker antigen-retrieval and the MS110 antibody staining. To assess the impact of BRCA1 germline mutations on protein localization, we retrospectively tested 16 of the tumor specimens to determine whether they contained the common Ashkenazi Jewish founder mutations in BRCA1 (185delAG, 5382insC), and BRCA2 (6174delT). We also compared co-localization of BRCA1 and nucleolin in MCF7 cells (wild type) and a mutant BRCA1 cell line, HCC1937 (5382insC).Results: In FFPE tissue, with MS110 antibody staining, we frequently found reduced BRCA1 nuclear staining in breast tumor tissue compared to normal tissue, and less BRCA1 staining with higher histological grade in the tumors. However, in the frozen sections, BRCA1 antibody staining showed punctate, intra-nuclear granules in varying numbers of tumor, lactating, and normal cells. Two mutation carriers were identified and were confirmed by gene sequencing. We have also compared co-localization of BRCA1 and nucleolin in MCF7 cells (wild type) and a mutant BRCA1 cell line, HCC1937 (5382insC) and found altered sub-nuclear and nucleolar localization patterns consistent with a functional impact of the mutation on protein localization.Conclusions: The data presented here support a role for BRCA1 in the pathogenesis of sporadic and inherited breast cancers. The use of well-characterized reagents may lead to further insights into the function of BRCA1 and possibly the further development of targeted therapeutics.

Original languageEnglish (US)
Article number70
JournalCancer Cell International
Volume13
Issue number1
DOIs
StatePublished - Jul 15 2013

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Keywords

  • BRCA1 mutations
  • Breast cancer
  • Frozen section immunohistology
  • Nucleolar localization

ASJC Scopus subject areas

  • Genetics
  • Oncology
  • Cancer Research

Cite this

Tulchin, N., Ornstein, L., Dikman, S., Strauchen, J., Jaffer, S., Nagi, C., Bleiweiss, I., Kornreich, R., Edelmann, L., Brown, K., Bodian, C., Nair, V. D., Chambon, M., Woods, N. T., & Monteiro, A. N. A. (2013). Localization of BRCA1 protein in breast cancer tissue and cell lines with mutations. Cancer Cell International, 13(1), [70]. https://doi.org/10.1186/1475-2867-13-70