title: Identification of miR-34a-target interactions by a combined network based and experimental approach.
abstract: Circulating miRNAs have been associated with numerous human diseases. The lack of understanding the functional roles of blood-born miRNAs limits, however, largely their value as disease marker. In a systems biology analysis we identified miR-34a as strongly associated with pathogenesis. Genome-wide analysis of miRNAs in blood cell fractions highlighted miR-34a as most significantly up-regulated in CD3+ cells of lung cancer patients. By our in silico analysis members of the protein kinase C family (PKC) were indicated as miR-34a target genes. Using a luciferase assay, we confirmed binding of miR-34a-5p to target sequences within the 3'UTRs of five PKC family members. To verify the biological effect, we transfected HEK 293T and Jurkat cells with miR-34a-5p causing reduced endogenous protein levels of PKC isozymes. By combining bioinformatics approaches with experimental validation, we demonstrate that one of the most relevant disease associated miRNAs has the ability to control the expression of a gene family.
authors.affiliation: Institute of Human Genetics, Saarland University, 66421 Homburg, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Hart
authors.affiliation: Institute of Human Genetics, Saarland University, 66421 Homburg, Germany.
authors.forename: Stefanie
authors.initials: S
authors.lastname: Rheinheimer
authors.affiliation: Institute of Human Genetics, Saarland University, 66421 Homburg, Germany.
authors.forename: Petra
authors.initials: P
authors.lastname: Leidinger
authors.affiliation: Chair for Clinical Bioinformatics, Saarland University, 66123 Saarbrücken, Germany.
authors.forename: Christina
authors.initials: C
authors.lastname: Backes
authors.affiliation: Institute of Virology, Saarland University Medical School, 66421 Homburg, Germany.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Menegatti
authors.affiliation: Chair for Clinical Bioinformatics, Saarland University, 66123 Saarbrücken, Germany.
authors.forename: Tobias
authors.initials: T
authors.lastname: Fehlmann
authors.affiliation: Institute of Virology, Saarland University Medical School, 66421 Homburg, Germany.
authors.forename: Friedrich
authors.initials: F
authors.lastname: Grässer
authors.affiliation: Chair for Clinical Bioinformatics, Saarland University, 66123 Saarbrücken, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Keller
authors.affiliation: Institute of Human Genetics, Saarland University, 66421 Homburg, Germany.
authors.forename: Eckart
authors.initials: E
authors.lastname: Meese
mesh_terms: D017252:CD3 Complex / Q000378:metabolism; D005786:Gene Expression Regulation / Q000235:genetics*; D053263:Gene Regulatory Networks / Q000235:genetics*; D055106:Genome-Wide Association Study; D006801:Humans; D008175:Lung Neoplasms / Q000235:genetics; D035683:MicroRNAs / Q000378:metabolism*; D011493:Protein Kinase C / Q000096:biosynthesis* / Q000235:genetics; D049490:Systems Biology / Q000379:methods; D013601:T-Lymphocytes / Q000378:metabolism; D015854:Up-Regulation
chemical_list: D017252:CD3 Complex; C522425:MIRN34 microRNA, human; D035683:MicroRNAs; D011493:Protein Kinase C
keywords: PKC family; cancer; immune system; miR-34a; network analysis