Detection of Mitochondrial DNA Mutations in Mammary Epithelial Cells in Nipple Aspirate Fluid
Abstract
We have successfully developed a method to isolate free DNA from ductal lavage (DL) and nipple aspirate fluid (NAF), and to use the isolated DNA for mitochondrial DNA (mtDNA) mutation analysis and LOH studies in order to improve early detection of breast cancer. We evaluated 26 DL and six NAF samples from 14 women of known BRAC1 status (9 carriers and 5 non-carriers), with no clinical evidence of breast tumors. LOH studies at the BRAC1 locus were possible in 19/26 DL samples, and at the FHIT locus in 16/26 samples. In 4/9 mutation carriers we found LOH at the BRAC1 allele, and in 2 of these we also found LOH at the FHIT allele. In one of the mutation carriers with BRAC1 LOH, invasive breast cancer was subsequently detected, and the tumor showed the same LOH as the DL. In one of the true negatives, BRAC1 and FHIT LOH were detected. The mitochondrial studies were possible in all 26 DL samples and a somatic mutation at the D310 marker was found in 3/9 carriers, 2 of whom also had LOH were detected. The mitochondrial studies were possible in all 26 DL samples and a somatic mutation at the D310 marker was found in 3/9 carriers, 2 of whom also has LOH at the BRAC1 locus, and in none of the non-carriers, mtDNA mutation evaluation was possible in 4/6 NAF samples. The NAF and DL results were concordant. One NAF sample from a BRCA1 patient showed a mtDNA mutation. Our data demonstrated the feasibility of molecular studies using the free DNA present in the ductal fluid, while the intact cells can be used for cytologic studies.
Document Details
- Document Type
- Technical Report
- Publication Date
- Sep 01, 2004
- Accession Number
- ADA434094
Entities
People
- Bassem R. Haddad
Organizations
- Georgetown University