title: DNA methylation contributes to deregulation of 12 cancer-associated microRNAs and breast cancer progression.
abstract: The methylation of promoter CpG islands and the interaction between microRNAs (miRNAs) and messenger RNAs (mRNAs) of target genes are considered two crucial mechanisms for gene and pathway deregulation in malignant tumors. The aim of this study was to analyze the role of promoter methylation in altering the expression of 13 miRNAs that are associated with breast cancer (BC): miR-124, -125b, -127, -132, -137, -148a, -191, -193a, -203, -212, -34b, -375, -9. The role of methylation in the deregulation of these miRNAs has not been previously assessed in the representative set of BC samples. We used a set of 58 paired (tumor/normal) breast tissue samples and methylation-specific PCR to demonstrate significant aberrations in the methylation patterns of 9 miRNA genes. In particular, we observed hypermethylation of MIR-127, -132, and -193a, and hypomethylation of MIR-191 for the first time. Using quantitative PCR, we established a strong correlation between promoter methylation and expression levels for 12 miRNA genes (all except MIR-212); this finding demonstrates the functional importance of altered methylation patterns. We also performed a correlation analysis between expression levels of the 13 miRNAs and 5 cancer-associated genes, namely RASSF1(A), CHL1, APAF1, DAPK1, and BCL2, which were predicted as targets for these miRNAs, to investigate the impact of these miRNAs on these genes with key cellular functions in BC. Significant negative correlation was revealed for the following miRNA-mRNA pairs: miR-127-5p and DAPK1, miR-375 and RASSF1(A), and miR-124-3p and BCL2. Additionally, we also found a strong association between hypermethylation of MIR-127 and MIR-125b-1 and BC progression, particularly metastasis. Thus, our findings provide evidence for the significant role of methylation in the deregulation of 12 miRNA genes in BC, identify putative novel functional miRNA-mRNA pairs, and suggest MIR-127 and MIR-125b-1 hypermethylation to be potential biomarkers of BC metastasis.
authors.affiliation: Institute of General Pathology and Pathophysiology, 125315 Moscow, Russia.
authors.forename: Irina V
authors.initials: IV
authors.lastname: Pronina
authors.affiliation: Institute of General Pathology and Pathophysiology, 125315 Moscow, Russia.
authors.forename: Vitaly I
authors.initials: VI
authors.lastname: Loginov
authors.affiliation: Institute of General Pathology and Pathophysiology, 125315 Moscow, Russia.
authors.forename: Alexey M
authors.initials: AM
authors.lastname: Burdennyy
authors.affiliation: Institute of General Pathology and Pathophysiology, 125315 Moscow, Russia.
authors.forename: Marina V
authors.initials: MV
authors.lastname: Fridman
authors.affiliation: Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, 119991 Moscow, Russia.
authors.forename: Vera N
authors.initials: VN
authors.lastname: Senchenko
authors.affiliation: Blokhin Cancer Research Center, 115478 Moscow, Russia.
authors.forename: Tatiana P
authors.initials: TP
authors.lastname: Kazubskaya
authors.affiliation: Blokhin Cancer Research Center, 115478 Moscow, Russia.
authors.forename: Nikolay E
authors.initials: NE
authors.lastname: Kushlinskii
authors.affiliation: Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, 119991 Moscow, Russia.
authors.forename: Alexey A
authors.initials: AA
authors.lastname: Dmitriev
authors.affiliation: Institute of General Pathology and Pathophysiology, 125315 Moscow, Russia. Electronic address: eleonora10_45@mail.ru.
authors.forename: Eleonora A
authors.initials: EA
authors.lastname: Braga
mesh_terms: D014408:Biomarkers, Tumor / Q000235:genetics* / Q000378:metabolism; D001943:Breast Neoplasms / Q000175:diagnosis / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D018270:Carcinoma, Ductal, Breast / Q000175:diagnosis / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D018899:CpG Islands; D019175:DNA Methylation; D044127:Epigenesis, Genetic; D005260:Female; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D008207:Lymphatic Metastasis; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D009367:Neoplasm Staging; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism
chemical_list: D014408:Biomarkers, Tumor; C531888:MIRN125 microRNA, human; C547276:MIRN127 microRNA, human; D035683:MicroRNAs; D012333:RNA, Messenger
keywords: Breast cancer; Cancer-associated genes; Deregulation of gene expression; Epigenetics; Methylation of promoter CpG islands; MicroRNA-mRNA pairs