title: Cell-free and cell-bound circulating DNA in breast tumours: DNA quantification and analysis of tumour-related gene methylation.
abstract: Tumour development is characterised by the increased circulating DNA (cirDNA) concentration and by tumour-related changes in blood plasma DNA. Concentration of cirDNA and methylation of RARbeta2, RASSF1A and HIC-1 gene promoters were investigated in cell-free and cell-surface-bound fractions from healthy donors, patients with breast cancer, and patients with breast fibroadenoma. Tumour development was shown to lead to significant changes in the distribution of cirDNA between cell-free and cell-surface-bound fractions. Analysis of RARbeta2 and RASSF1A methylation in the total cirDNA provides 95% diagnostic coverage in breast cancer patients, 60% in patients with benign lesions, and is without false-positive results in healthy women. Results of the study indicate that methylation-specific PCR of RARbeta2 and RASSF1A genes based on the total cirDNA combined with the quantitative analysis of cirDNA distribution between cell-bound and cell-free fractions in blood provide the sensitive and accurate detection and discrimination of malignant and benign breast tumours.
authors.affiliation: Institute of Chemical Biology and Fundamental Medicine, SD RAS, 8, Lavrentiev ave., Novosibirsk 630090, Russia.
authors.forename: T E
authors.initials: TE
authors.lastname: Skvortsova
authors.forename: E Y
authors.initials: EY
authors.lastname: Rykova
authors.forename: S N
authors.initials: SN
authors.lastname: Tamkovich
authors.forename: O E
authors.initials: OE
authors.lastname: Bryzgunova
authors.forename: A V
authors.initials: AV
authors.lastname: Starikov
authors.forename: N P
authors.initials: NP
authors.lastname: Kuznetsova
authors.forename: V V
authors.initials: VV
authors.lastname: Vlassov
authors.forename: P P
authors.initials: PP
authors.lastname: Laktionov
mesh_terms: D001943:Breast Neoplasms / Q000097:blood* / Q000235:genetics; D002474:Cell-Free System; D019175:DNA Methylation; D004273:DNA, Neoplasm / Q000097:blood*; D004268:DNA-Binding Proteins / Q000235:genetics*; D005260:Female; D018226:Fibroadenoma / Q000097:blood* / Q000235:genetics; D006801:Humans; D051741:Kruppel-Like Transcription Factors; D018168:Receptors, Retinoic Acid / Q000235:genetics*; D014157:Transcription Factors / Q000235:genetics*; D025521:Tumor Suppressor Proteins / Q000235:genetics*
chemical_list: D004273:DNA, Neoplasm; D004268:DNA-Binding Proteins; C499808:HIC1 protein, human; D051741:Kruppel-Like Transcription Factors; C411762:RASSF1 protein, human; D018168:Receptors, Retinoic Acid; D014157:Transcription Factors; D025521:Tumor Suppressor Proteins; C082925:retinoic acid receptor beta