RRC ID 71705
Author Shiraishi M, Chuu YH, Sekiya T.
Title Isolation of DNA fragments associated with methylated CpG islands in human adenocarcinomas of the lung using a methylated DNA binding column and denaturing gradient gel electrophoresis.
Journal Proc Natl Acad Sci U S A
Abstract We have constructed a library of DNA fragments heavily methylated in human adenocarcinomas of the lung to permit the comprehensive isolation of methylated CpG islands in cancer. Heavily methylated genomic DNA fragments from tumors of nine male patients were enriched using a methylated DNA binding column and used for construction of the library. From this library, DNA fragments having properties of CpG islands were isolated on the basis of their reduced rate of strand dissociation during denaturing gradient gel electrophoresis. Approximately 1,000 clones, corresponding to 0.3% of the library were analyzed, and nine DNA fragments were identified as being associated with CpG islands that were methylated in tumor DNA. One CpG island was methylated specifically in tumor DNA, whereas the remaining eight CpG islands were methylated both in normal and tumor DNA derived from the same patients. Our results suggest that the number of CpG islands methylated specifically in tumors is not large. The library, which contains DNA fragments from methylated CpG islands comprehensively, is expected to be valuable when elucidating epigenetic processes involved in carcinogenesis.
Volume 96(6)
Pages 2913-8
Published 1999-3-16
DOI 10.1073/pnas.96.6.2913
PMID 10077611
PMC PMC15869
MeSH Adenocarcinoma / genetics* Chromatography / methods* CpG Islands / genetics* DNA Methylation DNA, Neoplasm / genetics DNA, Neoplasm / isolation & purification* Electrophoresis / methods* Humans Lung Neoplasms / genetics* Male Molecular Sequence Data Polymerase Chain Reaction
IF 9.412
Human and Animal Cells A549(RCB0098) LC-2/ad(RCB0440)