Breast Cancer Research and Treatment

, Volume 152, Issue 2, pp 271–282 | Cite as

DNA repair capacity is impaired in healthy BRCA1 heterozygous mutation carriers

  • Tereza Vaclová
  • Gonzalo Gómez-López
  • Fernando Setién
  • José María García Bueno
  • José Antonio Macías
  • Alicia Barroso
  • Miguel Urioste
  • Manel Esteller
  • Javier Benítez
  • Ana Osorio
Preclinical study


BRCA1 germline mutations increase the lifetime risk of developing breast and ovarian cancers. However, taking into account the differences in disease manifestation among mutation carriers, it is probable that different BRCA1 mutations have distinct haploinsufficiency effects and lead to the formation of different phenotypes. Using lymphoblastoid cell lines derived from heterozygous BRCA1 mutation carriers and non-carriers, we investigated the haploinsufficiency effects of various mutation types using qPCR, immunofluorescence, and microarray technology. Lymphoblastoid cell lines carrying a truncating mutation showed significantly lower BRCA1 mRNA and protein levels and higher levels of gamma-H2AX than control cells or those harboring a missense mutation, indicating greater spontaneous DNA damage. Cells carrying either BRCA1 mutation type showed impaired RAD51 foci formation, suggesting defective repair in mutated cells. Moreover, compared to controls, cell lines carrying missense mutations displayed a more distinct expression profile than cells with truncating mutations, which is consistent with different mutations giving rise to distinct phenotypes. Alterations in the immune response pathway in cells harboring missense mutations point to possible mechanisms of breast cancer initiation in carriers of these mutations. Our findings offer insight into how various heterozygous mutations in BRCA1 could lead to impairment of BRCA1 function and provide strong evidence of haploinsufficiency in BRCA1 mutation carriers.


BRCA1 Haploinsufficiency DNA repair Gene expression profiling Breast cancer 



We thank the Confocal Microscopy Core Unit at CNIO for their help with high-throughput microscopy, and all the families who volunteered to participate in this study. The study was funded by the Spanish Ministry of Economy and Competitiveness (MINECO) SAF2010-20493 and the Spanish Network on Rare Diseases (CIBERER).

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

10549_2015_3459_MOESM1_ESM.docx (23 kb)
Supplementary material 1 (DOCX 22 kb)
10549_2015_3459_MOESM2_ESM.docx (1.6 mb)
Supplementary material 2 (DOCX 1683 kb)


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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Tereza Vaclová
    • 1
  • Gonzalo Gómez-López
    • 2
  • Fernando Setién
    • 3
  • José María García Bueno
    • 4
  • José Antonio Macías
    • 5
  • Alicia Barroso
    • 1
  • Miguel Urioste
    • 6
  • Manel Esteller
    • 3
    • 7
    • 8
  • Javier Benítez
    • 1
    • 9
    • 10
  • Ana Osorio
    • 1
    • 9
  1. 1.Human Genetics Group, Human Cancer Genetics ProgrammeSpanish National Cancer Research Centre (CNIO)MadridSpain
  2. 2.Bioinformatics Unit, Structural Biology and Biocomputing ProgrammeSpanish National Cancer Research Centre (CNIO)MadridSpain
  3. 3.Cancer Epigenetics Group, Cancer Epigenetics and Biology Program (PEBC)Bellvitge Biomedical Biomedical Research Institute (IDIBELL)BarcelonaSpain
  4. 4.Medical Oncology SectionComplejo Hospitalario Universitario de AlbaceteAlbaceteSpain
  5. 5.Hereditary Cancer Unit, Medical Oncology ServiceHospital Morales MeseguerMurciaSpain
  6. 6.Familial Cancer Unit, Human Cancer Genetics ProgrammeSpanish National Cancer Research Centre (CNIO)MadridSpain
  7. 7.Department of Physiological Sciences II, School of MedicineUniversity of BarcelonaBarcelonaSpain
  8. 8.Institucio Catalana de Recerca i Estudis Avançats (ICREA)BarcelonaSpain
  9. 9.Spanish Network on Rare Diseases (CIBERER)MadridSpain
  10. 10.Genotyping Unit (CEGEN), Human Cancer Genetics ProgrammeSpanish National Cancer Research Centre (CNIO)MadridSpain

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