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Assays for Hypermethylation of the BRCA1 Gene Promoter in Tumor Cells to Predict Sensitivity to PARP-Inhibitor Therapy

  • Ilsiya Ibragimova
  • Paul Cairns
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 780)

Abstract

The breast cancer 1 and 2, early onset (BRCA1 and BRCA2) genes are important for double-strand break repair by homologous recombination. Cells with inactivating mutations of the BRCA1 or BRCA2 tumor suppressor genes show increased sensitivity to Poly-ADP ribose polymerase (PARP)-inhibitors in vitro. Sporadic breast tumors with BRCA1 promoter hypermethylation show a similar phenotype to familial BRCA1 patient tumors termed “BRCAness.” Sporadic ovarian tumors with functional inactivation of BRCA1 by hypermethylation will also have the BRCA-deficiency phenocopy. The loss of BRCA1 expression associated with promoter hypermethylation will disrupt BRCA-associated DNA repair and may sensitize tumors to BRCA-directed therapies. Thus, the determination of methylation status of BRCA1 may be an important predictive classifier of response to PARP-inhibitor therapy. The methylation, and thereby functional, status of other genes implicated in the wider BRCA/homologous recombination (HR) pathway may also be relevant to the prediction of response to PARP-inhibitor therapy. Here, we describe the four optimal technologies for assaying the promoter methylation status of BRCA1 and/or other genes.

Key words

BRCA1 Hypermethylation PARP Bisulfite sequencing Pyrosequencing Quantitative MSP Methylation beadchip 

Notes

Acknowledgment

This work was supported by the Ovarian Cancer SPORE at Fox Chase Cancer Center (P50 CA083638).

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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Ilsiya Ibragimova
    • 1
  • Paul Cairns
    • 1
  1. 1.Departments of Surgical Oncology and PathologyFox Chase Cancer CenterPhiladelphiaUSA

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