Identification of miR-34a-target interactions by a combined network based and experimental approach
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Martin Hart1, Stefanie Rheinheimer1, Petra Leidinger1, Christina Backes2, Jennifer Menegatti3, Tobias Fehlmann2, Friedrich Grässer3, Andreas Keller2, Eckart Meese1
1Institute of Human Genetics, Saarland University, 66421 Homburg, Germany
2Chair for Clinical Bioinformatics, Saarland University, 66123 Saarbrücken, Germany
3Institute of Virology, Saarland University Medical School, 66421 Homburg, Germany
Martin Hart, email: [email protected]
Keywords: network analysis, PKC family, miR-34a, cancer, immune system
Received: March 08, 2016 Accepted: April 16, 2016 Published: April 29, 2016
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.
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