Neural stem cells inhibit melanin production by activation of Wnt inhibitors

Insik Hwang, Ju Hwang Park, Hang Soo Park, Kyung Ah Choi, Ki Cheon Seol, Seung Ick Oh, Seong Man Kang, Sunghoi Hong

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Background: Melanin for skin pigmentation is synthesized from tyrosine via an enzymatic cascade that is controlled by tyrosinase (TYR), tyrosinase-related protein 1 (TRP1), and dopachrome tautomerase/tyrosinase related protein 2 (Dct/TRP2), which are the targets of microphthalmia-associated transcription factor (MITF). MITF is a master regulator of pigmentation and a target of β-catenin in Wnt/β-catenin signaling during melanocyte differentiation. Stem cells have been used in skin pigmentation studies, but the mechanisms were not determined for the conditioned medium (CM)-mediated effects. Objectives: In this study, the inhibition and mechanisms of melanin synthesis were elucidated in B16 melanoma cells and UV-B irradiated C57/BL-6 mice that were treated with human neural stem cell-conditioned medium (NSC-CM). Methods: B16-F10 melanoma cells (1.5×104cells/well) and the shaved dorsal skin of mice were pretreated with various amount (5, 10, 20, 50, and 100%) of NSC-CM. Melanin contents and TYR activity were measured by a Spectramax spectrophotometer. The expression of TYR, TRP1, Dct/TRP2, MITF, β-catenin and Wnt inhibitors were evaluated by RT-PCR and western blot. The dorsal skin samples were analyzed by immunofluorescence with various antibodies and compared with that control of tissues. Results: Marked decreases were evident in melanin content and TYR, TRP1, DCT/TRP2, MITF, and β-catenin expression in B16 cells and C57/BL-6 mice. NSC-CM negatively regulated Wnt/β-catenin signaling by decreasing the expression of β-catenin protein, which resulted from robust expression of Wnt inhibitors Dickkopf-1 (DKK1) and secreted frizzled-related protein 2 (sFRP2). Conclusions: These results demonstrate that NSC-CM suppresses melanin production in vitro and in vivo, suggesting that factors in NSC-CM may play an important role in deregulation of epidermal melanogenesis.

Original languageEnglish
Pages (from-to)274-283
Number of pages10
JournalJournal of Dermatological Science
Volume72
Issue number3
DOIs
Publication statusPublished - 2013 Dec 1

Fingerprint

Catenins
Neural Stem Cells
Melanins
Conditioned Culture Medium
Microphthalmia-Associated Transcription Factor
Stem cells
Monophenol Monooxygenase
Chemical activation
Skin
Skin Pigmentation
Experimental Melanomas
Deregulation
Melanocytes
Spectrophotometers
Pigmentation
Fluorescent Antibody Technique
Tyrosine
Stem Cells
Western Blotting
Tissue

Keywords

  • Melanin
  • MITF
  • Neural stem cell-conditioned medium
  • Tyrosinase
  • Wnt inhibitors

ASJC Scopus subject areas

  • Dermatology
  • Biochemistry
  • Molecular Biology

Cite this

Neural stem cells inhibit melanin production by activation of Wnt inhibitors. / Hwang, Insik; Park, Ju Hwang; Park, Hang Soo; Choi, Kyung Ah; Seol, Ki Cheon; Oh, Seung Ick; Kang, Seong Man; Hong, Sunghoi.

In: Journal of Dermatological Science, Vol. 72, No. 3, 01.12.2013, p. 274-283.

Research output: Contribution to journalArticle

Hwang, Insik ; Park, Ju Hwang ; Park, Hang Soo ; Choi, Kyung Ah ; Seol, Ki Cheon ; Oh, Seung Ick ; Kang, Seong Man ; Hong, Sunghoi. / Neural stem cells inhibit melanin production by activation of Wnt inhibitors. In: Journal of Dermatological Science. 2013 ; Vol. 72, No. 3. pp. 274-283.
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AU - Choi, Kyung Ah

AU - Seol, Ki Cheon

AU - Oh, Seung Ick

AU - Kang, Seong Man

AU - Hong, Sunghoi

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N2 - Background: Melanin for skin pigmentation is synthesized from tyrosine via an enzymatic cascade that is controlled by tyrosinase (TYR), tyrosinase-related protein 1 (TRP1), and dopachrome tautomerase/tyrosinase related protein 2 (Dct/TRP2), which are the targets of microphthalmia-associated transcription factor (MITF). MITF is a master regulator of pigmentation and a target of β-catenin in Wnt/β-catenin signaling during melanocyte differentiation. Stem cells have been used in skin pigmentation studies, but the mechanisms were not determined for the conditioned medium (CM)-mediated effects. Objectives: In this study, the inhibition and mechanisms of melanin synthesis were elucidated in B16 melanoma cells and UV-B irradiated C57/BL-6 mice that were treated with human neural stem cell-conditioned medium (NSC-CM). Methods: B16-F10 melanoma cells (1.5×104cells/well) and the shaved dorsal skin of mice were pretreated with various amount (5, 10, 20, 50, and 100%) of NSC-CM. Melanin contents and TYR activity were measured by a Spectramax spectrophotometer. The expression of TYR, TRP1, Dct/TRP2, MITF, β-catenin and Wnt inhibitors were evaluated by RT-PCR and western blot. The dorsal skin samples were analyzed by immunofluorescence with various antibodies and compared with that control of tissues. Results: Marked decreases were evident in melanin content and TYR, TRP1, DCT/TRP2, MITF, and β-catenin expression in B16 cells and C57/BL-6 mice. NSC-CM negatively regulated Wnt/β-catenin signaling by decreasing the expression of β-catenin protein, which resulted from robust expression of Wnt inhibitors Dickkopf-1 (DKK1) and secreted frizzled-related protein 2 (sFRP2). Conclusions: These results demonstrate that NSC-CM suppresses melanin production in vitro and in vivo, suggesting that factors in NSC-CM may play an important role in deregulation of epidermal melanogenesis.

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