UVB-inhibited H19 activates melanogenesis by paracrine effects

Exp Dermatol. 2018 Oct;27(10):1120-1125. doi: 10.1111/exd.13749. Epub 2018 Aug 20.

Abstract

The long noncoding RNA H19 was reported to associate with melanogenesis. However, it remains unknown whether H19 expression will be changed by UVB irradiation and whether H19 will regulate melanocytes melanogenesis by paracrine effects. Here, we analysed the expression changes of H19 irradiated by UVB in keratinocytes and explored the mechanism of melanogenesis stimulated by H19 through paracrine effects. First, after keratinocytes were exposed to UVB irradiation, expression of H19 and pro-opiomelanocortin (POMC) was measured by qRT-PCR. Also, α-melanocyte-stimulating hormone (α-MSH) contents in cells supernatant were measured by ELISA. Then, H19 siRNAs were designed and transfected into keratinocytes by liposome. The expression changes of H19, POMC and α-MSH were detected. Besides, expression of p53 was detected by Western blot. After that, supernatant of keratinocytes with H19 siRNAs or negative control siRNA was cocultured with immortalized melanocyte line PIG1. Expression levels of MiTF, TYR, Rab27A, TYRP2, FSCN1 and MYO5A in PIG1 cells were detected by Western blot and qRT-PCR. We found that H19 expression of keratinocytes cells decreased after UVB irradiation. However, the levels of POMC, α-MSH and p53 were upregulated in UVB-irradiated cells. Compared with the negative control, H19 siRNAs could significantly increase the expression of POMC, α-MSH and p53. After supernatant of keratinocytes transfected with H19 siRNAs was cocultured with PIG1 cells, the levels of MiTF, TYR and Rab27A were upregulated in PIG1 cells. In conclusion, UVB-inhibited H19 may promote α-MSH secretion by p53 in keratinocytes and then regulate melanocytes melanogenesis through paracrine effects.

Keywords: UVB; H19; melanogenesis; p53; α-MSH.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Carrier Proteins / genetics
  • Carrier Proteins / metabolism
  • Cell Line
  • Cell Survival / radiation effects
  • Coculture Techniques
  • Cyclooxygenase 2 / genetics
  • Dose-Response Relationship, Radiation
  • Down-Regulation / radiation effects
  • Fibroblast Growth Factor 2 / genetics
  • Humans
  • Intramolecular Oxidoreductases / genetics
  • Intramolecular Oxidoreductases / metabolism
  • Keratinocytes / physiology
  • Keratinocytes / radiation effects
  • Melanins / biosynthesis*
  • Melanocytes / physiology
  • Melanocytes / radiation effects
  • Microfilament Proteins / genetics
  • Microfilament Proteins / metabolism
  • Microphthalmia-Associated Transcription Factor / genetics
  • Microphthalmia-Associated Transcription Factor / metabolism
  • Myosin Heavy Chains / genetics
  • Myosin Heavy Chains / metabolism
  • Myosin Type V / genetics
  • Myosin Type V / metabolism
  • Paracrine Communication / radiation effects*
  • Pro-Opiomelanocortin / genetics*
  • RNA, Long Noncoding / genetics*
  • RNA, Long Noncoding / metabolism*
  • RNA, Small Interfering / genetics
  • Tumor Suppressor Protein p53 / metabolism
  • Tyrosine / genetics
  • Tyrosine / metabolism
  • Ultraviolet Rays*
  • Up-Regulation / radiation effects
  • alpha-MSH / metabolism
  • rab27 GTP-Binding Proteins / genetics
  • rab27 GTP-Binding Proteins / metabolism

Substances

  • Carrier Proteins
  • FSCN1 protein, human
  • H19 long non-coding RNA
  • MITF protein, human
  • Melanins
  • Microfilament Proteins
  • Microphthalmia-Associated Transcription Factor
  • RNA, Long Noncoding
  • RNA, Small Interfering
  • Tumor Suppressor Protein p53
  • rab27 GTP-Binding Proteins
  • Fibroblast Growth Factor 2
  • MYO5A protein, human
  • Tyrosine
  • alpha-MSH
  • Pro-Opiomelanocortin
  • Cyclooxygenase 2
  • PTGS2 protein, human
  • Myosin Type V
  • RAB27A protein, human
  • Myosin Heavy Chains
  • Intramolecular Oxidoreductases
  • dopachrome isomerase