Identification of DNA methylation markers for human breast carcinomas using the methylation-sensitive restriction fingerprinting technique

Tim Hui Ming Huang, Douglas E. Laux, Brian C. Hamlin, Phuong Tran, Hoa Tran, Dennis B. Lubahn

Research output: Contribution to journalArticle

115 Scopus citations

Abstract

We have developed a PCR-based method, called methylation-sensitive restriction fingerprinting (MSRF), to screen changes in DNA methylation in breast carcinoma. Two hypermethylation-containing fragments, HBC-1 (for 'hypermethylation in breast cancer') and HBC-2, were identified in the amplified breast tumor DNA relative to the amplified normal breast DNA of a patient. Nucleotide sequence analysis revealed no significant matches between the sequence of HBC-1 and the known sequences in the GenBank database, whereas the sequence of HBC-2 matched the upstream region of an antisense WTI (Wilms' tumor suppressor gene) promoter. The methylation status in the breast tumor DNA from this patient was confirmed by Southern hybridization using HBC-1 and HBC-2 as probes, respectively. Further analysis showed that HBC-1 was methylated aberrantly in 90% (17 of 19 patients) of the primary breast carcinomas examined. This study demonstrates that MSRF provides a useful means for screening aberrant changes in DNA methylation during tumorigenesis. The commonly methylated fragments identified by MSRF could potentially supplement pathological markers currently used for cancers and additionally lead to the discovery of novel methylated tumor suppressor genes.

Original languageEnglish (US)
Pages (from-to)1030-1034
Number of pages5
JournalCancer Research
Volume57
Issue number6
StatePublished - Apr 9 1997
Externally publishedYes

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

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