Demethylation of a repetitive DNA sequence in human cancers

Electrophoresis. 1995 Feb;16(2):227-32. doi: 10.1002/elps.1150160138.

Abstract

To detect DNA alterations in unknown regions in human cancers, we have performed restriction landmark genomic scanning (RLGS) analysis of DNA isolated from cancer and normal cells. One spot with a highly intensified signal was detected in DNA from all six malignant melanoma cell lines, two of five colon cancer cell lines and one of six pancreatic cancer cell lines analyzed. In DNA from normal cells, two placentas and seven cultured lymphocytes, the signal of this spot was not intense. The DNA fragment corresponding to the spot was cloned. By nucleotide sequence analysis, the DNA fragment was revealed to be a part of a repeating unit of a 13 kbp nucleotide sequence of which 200 copies were located in chromosome 8q21. Southern blotting analysis using the cloned fragment as a probe demonstrated that the intensified signal for the DNA fragment observed in cancer cells was due to demethylation in the recognition sequence of the NotI restriction enzyme. The results suggest that marked demethylation in the repeating units might be associated with the genesis or progression of some types of cancers.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Blotting, Southern
  • Cloning, Molecular
  • Colonic Neoplasms / chemistry
  • Colonic Neoplasms / genetics
  • Colonic Neoplasms / metabolism
  • DNA, Neoplasm / chemistry
  • DNA, Neoplasm / genetics*
  • DNA, Neoplasm / metabolism
  • Deoxyribonucleases, Type II Site-Specific
  • Electrophoresis / methods*
  • Female
  • Genetic Techniques
  • Humans
  • Melanoma / chemistry
  • Melanoma / genetics
  • Melanoma / metabolism
  • Methylation
  • Molecular Sequence Data
  • Neoplasms / chemistry
  • Neoplasms / genetics*
  • Neoplasms / metabolism
  • Pancreatic Neoplasms / chemistry
  • Pancreatic Neoplasms / genetics
  • Pancreatic Neoplasms / metabolism
  • Placenta / metabolism
  • Pregnancy
  • Repetitive Sequences, Nucleic Acid*
  • Tumor Cells, Cultured

Substances

  • DNA, Neoplasm
  • Deoxyribonucleases, Type II Site-Specific
  • GCGGCCGC-specific type II deoxyribonucleases

Associated data

  • GENBANK/L09552