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Aldosterone‐regulated miRNAs and their target genes in mouse cortical collecting duct mpkCCD cells
Author(s) -
Kwon TaeHwan,
Park EuiJung,
Jung Hyun Jun,
Choi HyoJung
Publication year - 2017
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.31.1_supplement.855.16
Subject(s) - microrna , biology , kegg , wnt signaling pathway , gene , microarray , fold change , gene expression profiling , dna microarray , microarray analysis techniques , gene expression , computational biology , microbiology and biotechnology , transcriptome , genetics
Background Mature microRNA (miRNA) is a modulator in the post‐transcriptional regulation. The present study aimed 1) to profile the aldosterone‐regulated miRNAs in mouse cortical collecting duct cells (mpkCCDc14 cells); 2) to identify the main signaling pathways where putative target genes of identified miRNAs are enriched; and 3) to determine the roles of selected miRNAs and target genes in aldosterone‐mediated action. Methods Microarray chip assay (Affymetrix GeneChip miRNA 4.0 array) was done in the mpkCCDc14 cells in the absence or the presence of aldosterone treatment (10 −6 M) for 3 d. The candidate miRNAs were selected by 1) more or less than 30% of significant fold‐changes (Protocol 1), or 2) differential expression analysis carried out using the R package ‘bridge’ (Bayesian Robust Inference for Differential Gene Expression. R package version 1.34.0., Protocol 2). Both the putative target genes of identified miRNAs and miRNAs‐enriched pathways were analyzed by using DIANA‐mirPath program. Results Twenty‐nine miRNAs were significantly up‐regulated (> 1.3‐fold) and 27 miRNAs were markedly down‐regulated (< 0.7‐fold) in mpkCCDc14 cells after aldosterone treatment (Protocol 1). Moreover, 5 up‐regulated and 7 down‐regulated miRNAs (> 1.2‐fold and < 0.8‐fold) were selected with high posterior probabilities (> 0.95, Protocol 2). Putative target genes of identified miRNAs were associated with 55 KEGG pathways (Protocol 1) and 29 KEGG pathways (Protocol 2) by DIANA‐mirPath program analysis. Among them, Wnt signaling pathway was particularly the most highly ranked. Seven up‐regulated and 8 down‐regulated mature miRNAs were enriched in the Wnt signaling pathways. Their quantitative changes were further examined by qPCR, revealing that miR‐130b‐3p, miR‐34c‐5p and miR‐146a‐5p were significantly changed. Six target genes ( Rock1, skp1a, Tbl1xr1, Ppp2re, Wnt2b, Plcb1 ) of miR‐130b‐3p, 6 target genes ( Lef1, Camk2b, Tbl1xr1, Prickle1, Ppp2r5a, Daam1 ) of miR‐34c‐5p and 4 target genes ( Ppp3r2, Cxxc4, Smad4, Nfat5 ) of miR‐146a‐5p were identified. Conclusion Aldosterone induces significant changes of miRNAs expression in mpkCCDc14 cells, and Wnt signaling is the most highly ranked pathway where identified miRNAs are enriched. The changes of target genes and corresponding protein expression may potentially be involved in the aldosterone‐mediated renal fibrosis, which need to be further examined.