DNA Methylation Alterations in Breast Cancer
Abstract
We proposed to investigate breast cancer-associated DNA methylation alterations using a technique named Methylation Sensitive Restriction Fragment Length Polymorphism (MS-AFLP) which we developed. We performed NotI-MseI MS-AFLP using clinical specimens of normal and tumor breast DNA. We used both combinations of four NotI and four MseI primers with an additional selective residue at the 3' end (4x4 format) and combinations of four NotI with one additional residue and sixteen MseI with two additional selective residues (4x16 format). We have identified many bands that exhibited alterations in band intensity between normal and tumor breast DNA fingerprints. We have cut out the dried gel slices from those bands, eluted and PCR-amplified DNA fragments, and cloned into a plasmid vector. After transformation of E. coli bacteria, the nucleotide sequences of inserts were determined from several transformant clones. For some of the interesting bands, we have also transferred DNA from the DNA fingerprints by electroblotting onto nylon membranes, hybridized those membranes with fragment probes prepared from individual clones, and determined the identity of the fragments showing alterations. From these studies, we have obtained results indicating the possible associations of homeotic genes and related genes in the pathogenesis of breast cancer.
Document Details
- Document Type
- Technical Report
- Publication Date
- Jul 01, 2000
- Accession Number
- ADA385623
Entities
People
- Fumiichiro Yamamoto
Organizations
- Sanford Burnham Prebys Medical Discovery Institute