Regular Article

British Journal of Cancer (2001) 84, 482–488. doi:10.1054/bjoc.2000.1627 www.bjcancer.com
Published online 13 February 2001

Screening of male breast cancer and of breast-ovarian cancer families for BRCA2 mutations using large bifluorescent amplicons

S Pages1, V Caux1, D Stoppa-Lyonnet1 and M Tosi2,3

  1. 1Service de Génétique Oncologique, Institut Curie, 26 rue d'Ulm 75248, Paris Cedex 5
  2. 2Unité d'Immunogénétique et INSERM U 276, Institut Pasteur, 25 rue du Docteur Roux, Paris, 75724, Cedex 15
  3. 3INSERM EMI 9906, Faculté de Médecine et Pharmacie, 76183 Rouen, France

Correspondence: M Tosi, INSERM EH19906, Faculté de Médecine et Pharmacie, 76183 Roven, France

Received 24 July 2000; Revised 18 October 2000; Accepted 8 November 2000.

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Abstract

41 breast cancer or breast-ovarian cancer families, including 12 families with at least one affected first-degree male relative, were screened for mutations in the BRCA2 gene. Mutations had not been found in the BRCA1 gene of these families. Chemical cleavage of Mismatch was used to identify nucleotide changes within large PCR products (average size 1.2 kb) that carried strand-specific fluorescent end-labels. 15 amplicons were sufficient to scan 18 exons, including the large exon 11. The remaining 9 small exons were examined by Denaturing Gradient Gel Electrophoresis. The high sensitivity of this approach was documented by the detection, in these 41 patients, of all 9 exonic single nucleotide polymorphisms reported with heterozygosity >0.1. Truncating BRCA2 mutations were found in 7 of the 41 families. 3 of them were in the group of 12 families comprising cases of male breast cancer. Since the methods used here have no bias for particular types of mutations, these data confirm the high proportion of frameshifts among mutations in BRCA2. However, relevant single nucleotide substitutions were also found: one resulting in a stop codon and another one, present in a male patient, was the previously reported change Asp2723His, that affects a highly conserved region of the BRCA2 protein. This study indicates a BRCA2 contribution of 10% (95% CI 2.5–17.5) to our original cohort of 59 breast-ovarian cancer families, whereas the contribution of BRCA1 had been estimated at 46% (95% CI 33–59). © 2001 Cancer Research Campaign

Keywords:

male breast cancer, breast-ovarian cancer, FAMA, chemical cleavage of mismatch, chimeric PCR primers

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