Skip to Content
Merck
CN
  • Stem Cell Factor-Inducible MITF-M Expression in Therapeutics for Acquired Skin Hyperpigmentation.

Stem Cell Factor-Inducible MITF-M Expression in Therapeutics for Acquired Skin Hyperpigmentation.

Theranostics (2020-01-07)
Cheong-Yong Yun, Eunmiri Roh, Song-Hee Kim, Jinhe Han, Jiyeon Lee, Da-Eun Jung, Ga Hyeon Kim, Sang-Hun Jung, Won-Jea Cho, Sang-Bae Han, Youngsoo Kim
ABSTRACT

Rationale: Microphthalmia-associated transcription factor M (MITF-M) plays important roles in the pigment production, differentiation and survival of melanocytes. Stem cell factor (SCF) and its receptor KIT stimulate MITF-M activity via phosphorylation at the post-translation level. However, the phosphorylation shortens half-life of MITF-M protein over the course of minutes. Here, we investigated novel hypotheses of (i) whether SCF/KIT can regulate MITF-M activity through gene expression as the alternative process, and (ii) whether chemical inhibition of KIT activity can mitigate the acquired pigmentation in skin by targeting the expression of MITF-M. Methods: We employed melanocyte cultures in vitro and pigmented skin samples in vivo, and applied immunoblotting, RT-PCR, siRNA-based gene knockdown and confocal microscopy. Results: The protein and mRNA levels of MITF-M in epidermal melanocytes and the promoter activity of MITF-M in B16-F0 melanoma cells demonstrated that SCF/KIT could trigger the expression of MITF-M de novo, following the phosphorylation-dependent proteolysis of pre-existing MITF-M protein. SCF/KIT regulated the transcription abilities of cAMP-responsive element-binding protein (CREB), CREB-regulated co-activator 1 (CRTC1) and SRY-related HMG-box 10 (SOX10) but not β-catenin at the MITF-M promoter. Meanwhile, chemical inhibition of KIT activity abolished SCF-induced melanin production in epidermal melanocyte cultures, as well as protected the skin from UV-B-induced hyperpigmentation in HRM2 mice or brownish guinea pigs, in which it down-regulated the expression of MITF-M de novo at the promoter level. Conclusion: We propose the targeting of SCF/KIT-inducible MITF-M expression as a strategy in the therapeutics for acquired pigmentary disorders.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
SU 6656, ≥98% (HPLC)
Sigma-Aldrich
SB 202190, ≥98% (HPLC)
Sigma-Aldrich
3,3′,5,5′-Tetramethylbenzidine, ≥99%
Sigma-Aldrich
ISCK03, ≥98% (HPLC), solid
Sigma-Aldrich
BIO, ≥98% (HPLC)
Sigma-Aldrich
Thiazolyl Blue Tetrazolium Bromide, powder, BioReagent, suitable for cell culture, suitable for insect cell culture, ≥97.5% (HPLC)
Sigma-Aldrich
Formaldehyde solution, for molecular biology, 36.5-38% in H2O
Sigma-Aldrich
TWEEN® 20, viscous liquid
Sigma-Aldrich
HEPES, BioPerformance Certified, ≥99.5% (titration), suitable for cell culture
Sigma-Aldrich
Sodium orthovanadate, ≥90% (titration)
Sigma-Aldrich
Rp-Adenosine 3′,5′-cyclic monophosphorothioate triethylammonium salt, powder, ≥98% (HPLC)
Sigma-Aldrich
U0126 ethanolate, ≥98% (HPLC), powder
Sigma-Aldrich
SB 216763, >98% (HPLC)
Sigma-Aldrich
Chromatin Immunoprecipitation (ChIP) Assay Kit, Contains all necessary reagents to perform 22 individual chromatin immunoprecipitation (ChIP) reactions using inexpensive protein A agarose beads.
Sigma-Aldrich
Imatinib
Sigma-Aldrich
PP2, ≥98% (HPLC)
Sigma-Aldrich
H-89 dihydrochloride hydrate, ≥98% (HPLC), powder
Sigma-Aldrich
LY-294,002 hydrochloride, solid, ≥98% (HPLC)
Sigma-Aldrich
PD 98,059, solid
Sigma-Aldrich
SB 203580, solid, ≥98% (HPLC)
Sigma-Aldrich
Arbutin, ≥98% (HPLC)
Sigma-Aldrich
Paraffin wax, mp 58-62 °C (ASTM D 87)
Sigma-Aldrich
Imatinib mesylate, ≥98% (HPLC)
Sigma-Aldrich
Dimethyl sulfoxide, suitable for HPLC, ≥99.7%
Sigma-Aldrich
Gelatin from cold water fish skin, solid
Sigma-Aldrich
Triton X-100, laboratory grade
Sigma-Aldrich
ECO TWEEN® 20, viscous liquid