Open AccessIdentification of miR-34a-target interactions by a combined network based and experimental approach
Author(s) -
Martin Hart,
Stefanie Rheinheimer,
Petra Leidinger,
Christina Backes,
Jennifer Menegatti,
Tobias Fehlmann,
Friedrich A. Grässer,
Andreas Keller,
Eckart Meese
Publication year - 2016
Publication title -
oncotarget
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.373
H-Index - 127
ISSN - 1949-2553
DOI - 10.18632/oncotarget.9103
Subject(s) - in silico , microrna , transfection , hek 293 cells , jurkat cells , biology , gene , computational biology , protein kinase c , luciferase , isozyme , pathogenesis , bioinformatics , kinase , microbiology and biotechnology , genetics , immunology , enzyme , t cell , biochemistry , immune system
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.