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- Title
Reduced MiR-675 in Exosome in H19 RNA-Related Melanogenesis via MITF as a Direct Target.
- Authors
Kim, Nan-Hyung; Choi, Soo-Hyun; Kim, Chang-Hyun; Lee, Chang Hoon; Lee, Tae Ryong; Lee, Ai-Young
- Abstract
H19 non-coding RNA downregulation stimulates melanogenesis in melasma patients. However, its mechanism is unclear. In this study, the potential role of a H19 microRNA, miR-675, in melanogenesis was examined. Real-time PCR using cultured normal human skin keratinocytes, melanocytes, and fibroblasts with or without H19 knockdown showed accompanying changes between expression levels of H19 and those of miR-675 in keratinocytes. MiR-675 was also detected in concentrated culture supernatants and showed expression levels parallel with those of cell lysates. In addition to RNase resistance, FACS analysis showed anti-CD63-positive exosomes in culture supernatants, suggesting miR-675 could be released extracellularly and delivered to neighboring cells without degradation. In western blot analysis, the miR-675 mimic reduced the expression of microphthalmia-associated transcription factor (MITF) and phosphorylation of cAMP-responsive element-binding protein, extracellular signal-regulated kinase and apoptosis signal-regulating kinase, whereas these expressions were increased by the miR-675 inhibitor. Although H19 was not a miR-675 target, luciferase reporter assay showed a direct binding of miR-675 to 3′-untranslated region of MITF. In addition, localized in vivo miR-675 overexpression in mouse using a cationic polymer transfection reagent showed reduced mRNA expression levels of MITF, tyrosinase, tyrosine-related protein-1 (Trp-1), and Trp-2. Collectively, the results suggest that miR-675 derived from keratinocytes could be involved in H19-stimulated melanogenesis using MITF as a target of miR-675.
- Subjects
NON-coding RNA; EXOSOMES; MELANOGENESIS; MELANOSIS; POLYMERASE chain reaction; KERATINOCYTES; FIBROBLASTS; MICROPHTHALMIA-associated transcription factor; APOPTOSIS
- Publication
Journal of Investigative Dermatology, 2014, Vol 134, Issue 4, p1075
- ISSN
0022-202X
- Publication type
Article
- DOI
10.1038/jid.2013.478