title: High-resolution melting-based quantitative analysis of RASSF1 methylation in breast cancer.
abstract: OBJECTIVE
  Breast cancer is the leading cause of death from cancer among women worldwide. The aberrant promoter methylation of tumor suppressor genes is a typical epigenetic alteration for breast cancer and can be detected in early carcinogenesis. High-throughput and cost-effective methods are needed for the early and sensitive detection of epigenetic changes in clinical material. The main purpose of our study was to optimize a high-resolution melting (HRM) assay for the reliable and quantitative assessment of RASSF1 gene methylation, which is considered one of the earliest epigenetic alterations in breast cancer.
  
  
  METHODS
  A total of 76 breast carcinomas and 10 noncancerous breast tissues were studied by means of HRM and compared with the results obtained by means of quantitative methylation-specific polymerase chain reaction (QMSP) and methylation-specific polymerase chain reaction (MSP).
  
  
  RESULTS
  Both quantitative methods, HRM and QMSP, showed a similar specificity and sensitivity for the detection of RASSF1 methylation in breast cancer (about 80% and 70%, respectively). In breast cancer, the mean methylation intensity of RASSF1 was 42.5% and 48.6% according to HRM and QMSP, respectively. Both methods detected low levels of methylation (less than 5%) in noncancerous breast tissues. In comparison with quantitative methods, MSP showed a lower sensitivity (70%), but a higher specificity (80%) for the detection of RASSF1 methylation in breast cancer.
  
  
  CONCLUSIONS
  HRM is as a simple, cost-effective method for the reliable high-throughput quantification of DNA methylation in clinical material.
authors.affiliation: Division of Human Genome Research Centre, Faculty of Natural Sciences, Vilnius University, M. K. Čiurlionio 21, 03101 Vilnius, Lithuania. sonata.jarmalaite@gf.vu.lt
authors.forename: Kristina
authors.initials: K
authors.lastname: Stuopelytė
authors.forename: Kristina
authors.initials: K
authors.lastname: Daniūnaitė
authors.forename: Aida
authors.initials: A
authors.lastname: Laurinavičienė
authors.forename: Valerijus
authors.initials: V
authors.lastname: Ostapenko
authors.forename: Sonata
authors.initials: S
authors.lastname: Jarmalaitė
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001943:Breast Neoplasms / Q000235:genetics*; D019175:DNA Methylation; D005260:Female; D006801:Humans; D008875:Middle Aged; D009691:Nucleic Acid Denaturation; D016133:Polymerase Chain Reaction / Q000379:methods*; D044366:Transition Temperature; D025521:Tumor Suppressor Proteins / Q000235:genetics*
chemical_list: C411762:RASSF1 protein, human; D025521:Tumor Suppressor Proteins