title: Identification of miR-199a-5p target genes in the skin keratinocyte and their expression in cutaneous squamous cell carcinoma.
abstract: BACKGROUND
  MicroRNAs (miRNAs) are small non-coding RNA molecules that mediate the biological cellular processes via regulation of target genes through translational repression or mRNA degradation. Among various miRNAs, miRNA-199a (miR-199a) has been known to be involved in cancer development and progression, protection of cardiomyocyte, and skeletal formation.
  
  
  OBJECTIVE
  Although miR-199a-5p was studied in various cell types, the role of miR-199a-5p and its target genes in skin keratinocyte have not been documented. In this study, we identified target genes of miR-199a-5p in skin keratinocyte.
  
  
  METHODS
  In order to identify the target of miR-199a-5p in keratinocyte, microarray analysis was performed. The relative expression of candidate target genes was investigated using quantitative RT-PCR and western blot analysis. To determine whether their expression was directly regulated by miR-199a-5p, luciferase reporter assay was performed. In order to investigate expression of target genes in cutaneous squamous cell carcinoma, immunohistochemistry was performed.
  
  
  RESULTS
  We identified new target genes, Bcam, Fzd6, and Wnt7a, as well as previously known targets, Ddr1 and Podxl. We found that their expressions were directly regulated by miR-199a-5p in the skin keratinocyte using in vitro study and observed that expression of miR-199a-5p was inversely correlated with those of BCAM, FZD6 and DDR1 in the cSCC. In addition, overexpression of miR-199a-5p resulted in inhibition of the migratory capability of the skin keratinocyte.
  
  
  CONCLUSIONS
  These results suggested that miR-199a-5p plays a role in pathogenesis of cSCC via inhibition of invasiveness through regulation of BCAM, FZD6 and DDR1 expression.
authors.affiliation: Department of Medical Life Sciences, The Catholic University of Korea, Banpodong 505, Seoul 137-701, Seochogu, South Korea.
authors.forename: Bong-Kyu
authors.initials: BK
authors.lastname: Kim
authors.affiliation: Department of Medical Life Sciences, The Catholic University of Korea, Banpodong 505, Seoul 137-701, Seochogu, South Korea.
authors.forename: Injung
authors.initials: I
authors.lastname: Kim
authors.affiliation: Department of Medical Life Sciences, The Catholic University of Korea, Banpodong 505, Seoul 137-701, Seochogu, South Korea. Electronic address: sjkyoon@catholic.ac.kr.
authors.forename: Sungjoo Kim
authors.initials: SK
authors.lastname: Yoon
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002294:Carcinoma, Squamous Cell / Q000235:genetics* / Q000378:metabolism; D002478:Cells, Cultured; D051157:Frizzled Receptors / Q000378:metabolism; D015870:Gene Expression / Q000502:physiology; D006801:Humans; D017403:In Situ Hybridization; D015603:Keratinocytes / Q000378:metabolism*; D051379:Mice; D035683:MicroRNAs / Q000235:genetics*; D012878:Skin Neoplasms / Q000235:genetics* / Q000378:metabolism; D051153:Wnt Proteins / Q000378:metabolism
chemical_list: D051157:Frizzled Receptors; C111048:Fzd6 protein, mouse; D035683:MicroRNAs; D051153:Wnt Proteins; C490508:Wnt7a protein, mouse; C531408:mirn199 microRNA, human
keywords: Bcam; Ddr1; Fzd6; Keratinocyte; MiR-199a-5p; Podxl; Wnt7a; cSCC