title: Identification of microRNA-mRNA functional interactions in UVB-induced senescence of human diploid fibroblasts.
abstract: BACKGROUND
  Cellular senescence can be induced by a variety of extrinsic stimuli, and sustained exposure to sunlight is a key factor in photoaging of the skin. Accordingly, irradiation of skin fibroblasts by UVB light triggers cellular senescence, which is thought to contribute to extrinsic skin aging, although molecular mechanisms are incompletely understood. Here, we addressed molecular mechanisms underlying UVB induced senescence of human diploid fibroblasts.
  
  
  RESULTS
  We observed a parallel activation of the p53/p21(WAF1) and p16(INK4a)/pRb pathways. Using genome-wide transcriptome analysis, we identified a transcriptional signature of UVB-induced senescence that was conserved in three independent strains of human diploid fibroblasts (HDF) from skin. In parallel, a comprehensive screen for microRNAs regulated during UVB-induced senescence was performed which identified five microRNAs that are significantly regulated during the process. Bioinformatic analysis of miRNA-mRNA networks was performed to identify new functional mRNA targets with high confidence for miR-15a, miR-20a, miR-20b, miR-93, and miR-101. Already known targets of these miRNAs were identified in each case, validating the approach. Several new targets were identified for all of these miRNAs, with the potential to provide new insight in the process of UVB-induced senescence at a genome-wide level. Subsequent analysis was focused on miR-101 and its putative target gene Ezh2. We confirmed that Ezh2 is regulated by miR-101 in human fibroblasts, and found that both overexpression of miR-101 and downregulation of Ezh2 independently induce senescence in the absence of UVB irradiation. However, the downregulation of miR-101 was not sufficient to block the phenotype of UVB-induced senescence, suggesting that other UVB-induced processes induce the senescence response in a pathway redundant with upregulation of miR-101.
  
  
  CONCLUSIONS
  We performed a comprehensive screen for UVB-regulated microRNAs in human diploid fibroblasts, and identified a network of miRNA-mRNA interactions mediating UVB-induced senescence. In addition, miR-101 and Ezh2 were identified as key players in UVB-induced senescence of HDF.
authors.affiliation: Institute for Biomedical Aging Research, Austrian Academy of Sciences, Rennweg 10, Innsbruck 6020, Austria. Pidder.Jansen-Duerr@uibk.ac.at.
authors.forename: Ruth
authors.initials: R
authors.lastname: Greussing
authors.forename: Matthias
authors.initials: M
authors.lastname: Hackl
authors.forename: Pornpimol
authors.initials: P
authors.lastname: Charoentong
authors.forename: Alexander
authors.initials: A
authors.lastname: Pauck
authors.forename: Rossella
authors.initials: R
authors.lastname: Monteforte
authors.forename: Maria
authors.initials: M
authors.lastname: Cavinato
authors.forename: Edith
authors.initials: E
authors.lastname: Hofer
authors.forename: Marcel
authors.initials: M
authors.lastname: Scheideler
authors.forename: Michael
authors.initials: M
authors.lastname: Neuhaus
authors.forename: Lucia
authors.initials: L
authors.lastname: Micutkova
authors.forename: Christoph
authors.initials: C
authors.lastname: Mueck
authors.forename: Zlatko
authors.initials: Z
authors.lastname: Trajanoski
authors.forename: Johannes
authors.initials: J
authors.lastname: Grillari
authors.forename: Pidder
authors.initials: P
authors.lastname: Jansen-Dürr
mesh_terms: D045744:Cell Line, Tumor; D016922:Cellular Senescence / Q000235:genetics* / Q000528:radiation effects; D004171:Diploidy; D000071221:Enhancer of Zeste Homolog 2 Protein; D005347:Fibroblasts / Q000378:metabolism* / Q000528:radiation effects; D053263:Gene Regulatory Networks / Q000528:radiation effects; D006801:Humans; D035683:MicroRNAs / Q000502:physiology*; D063151:Polycomb Repressive Complex 2 / Q000235:genetics* / Q000378:metabolism; D034622:RNA Interference / Q000528:radiation effects; D012333:RNA, Messenger / Q000235:genetics*; D059467:Transcriptome / Q000528:radiation effects; D014466:Ultraviolet Rays
chemical_list: C534072:MIRN101 microRNA, human; D035683:MicroRNAs; D012333:RNA, Messenger; C498409:EZH2 protein, human; D000071221:Enhancer of Zeste Homolog 2 Protein; D063151:Polycomb Repressive Complex 2
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