title: Trichostatin A induces a unique set of microRNAs including miR-129-5p that blocks cyclin-dependent kinase 6 expression and proliferation in H9c2 cardiac myocytes.
abstract: The pan-histone deacetylase inhibitor (HDACI), trichostatin A (TSA), was shown to normalize interleukin-18-induced cardiac hypertrophy in vivo and in vitro; evidently, this occurred via epigenetic mechanisms that profoundly altered cardiac gene expression (Majumdar et al. in, Physiol Genom, 43: 1392, 2011; BMC Genom, 13: 709, 2012). Here, we tested the hypothesis that TSA-induced changes in chromatin architecture also led to altered expression of microRNAs that in turn, contributed to the unique transcriptome of cardiac myocytes exposed to the HDACI. Using miRCURY LNA™ Universal microRNA PCR system, we demonstrate that H9c2 cells exposed to TSA for 6 and 24 h elicited differential expression of 19 and 16 microRNAs, respectively. H9c2 cells incubated in medium-containing 100 nM of TSA elicited a rapid and robust induction of miR-129-5p. Enhanced expression of miR-129-5p was also observed in the hearts of TSA-treated mice. Induction of miR-129-5p in H9c2 cells was accompanied by reduced expression of its direct target, cyclin-dependent kinase 6 (CDK6) that is a key regulator of cell cycle. Using cell division-dependent dilution of Cell Trace™ violet measurements we showed that concomitant induction of miR-129-5p and reduced CDK6 expression were mechanistically involved in TSA-induced inhibition of proliferation of H9c2 cells. Consistent with this scenario, cells expressing an antagomiR of miR-129-5p were resistant to the anti-proliferative actions of TSA. These data indicate that although TSA treatment led to altered expression of several microRNAs, the overarching action of TSA (i.e., inhibition of cell division) in H9c2 cells was achieved via miR-129-5p.
authors.affiliation: Department of Veterans Affairs Medical Center, Memphis, TN, 38104, USA.|Department of Pharmacology, University of Tennessee Health Science Center, Memphis, TN, 38163, USA.
authors.forename: Gipsy
authors.initials: G
authors.lastname: Majumdar
authors.affiliation: Department of Veterans Affairs Medical Center, Memphis, TN, 38104, USA. rraghow@uthsc.edu.|Department of Pharmacology, University of Tennessee Health Science Center, Memphis, TN, 38163, USA. rraghow@uthsc.edu.
authors.forename: Rajendra
authors.initials: R
authors.lastname: Raghow
mesh_terms: D000818:Animals; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects*; D051361:Cyclin-Dependent Kinase 6 / Q000037:antagonists & inhibitors*; D006801:Humans; D006877:Hydroxamic Acids / Q000494:pharmacology*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D035683:MicroRNAs / Q000235:genetics*; D032383:Myocytes, Cardiac / Q000187:drug effects* / Q000201:enzymology / Q000378:metabolism
chemical_list: D006877:Hydroxamic Acids; D035683:MicroRNAs; C541915:Mirn129 microRNA, human; C012589:trichostatin A; C495911:CDK6 protein, human; D051361:Cyclin-Dependent Kinase 6
keywords: H9c2 cardiac myocytes; HDAC inhibitor; TSA; microRNA