Sulforaphane, found widely in broccoli, inhibits melanin synthesis and tyrosinase expression in B16 mouse melanoma cells. The study found that 5M sulforaphane inhibited melanogenesis to an equivalent level as 100M arbutin. Sulforaphane activates phosphorylated ERK and inhibits phosphorylated p38, affecting the mitogen-activated protein kinase family which controls tyrosinase expression. By regulating the MAP kinase family, sulforaphane inhibits melanogenesis and tyrosinase without cytotoxic effects, indicating it may be an effective skin-whitening agent.

1. Sulforaphane Inhibited Melanin Synthesis
by Regulating Tyrosinase Gene Expression
in B16 Mouse Melanoma Cells
1
▶Objective : Sulforaphane과 Tyrosinase와 Melanogenesis에 대한 관계를
알아보기

2. Sulforaphane is a compound widely present in consumed vegetables, particularly broccoli.
Previous studies have demonstrated that sulforaphane has many physiological effects
including anti-cancer, anti-oxidation, and detoxification. In this study, we found that
sulforaphane inhibited melanogenesis and tyrosinase expression. The inhibitory effect of
5M sulforaphane on melanogenesis was determined to be equivalent to that of 100M arbutin.
In addition, sulforaphane induced phosphorylated extracellular signal-regulated kinase (ERK)
and inhibited phosphorylated p38. It has been reported that the phosphorylated
mitogenactivated protein (MAP) kinase family (ERK and p38) controls tyrosinase expression.
Our data indicate that sulforaphane inhibited melanogenesis and tyrosinase expression by
affecting the phosphorylated MAP kinase family. These findings indicate that sulforaphane
might be an effective skin-whitening agent.
2

3. 5M sulforaphane ≒ 100M arbutin
▶sulforaphane inhibits melanogenesis and tyrosinase expression.
▶MAP kinase
: mitogen-activated protein kinase
▶ERK
: extracellular signal-regulated kinase
sulforaphane
<activation> phosphorylated ERK
<inhibition> phosphorylated p38
tyrosinase expressionmelanin
ERK p38
MAP kinase family
3
<regulation>

4. sulforaphane
 in vegetable, broccoli, broccoli sprouts
 myrosinase : glucoraphanin  sulforaphane
 isothiocyanate group
 skin-whitening agent
 antioxidant, help remove ROS
 exhibit anti-cancer and antimicrobial properties
<출처 : wikipedia>
4

5. 5
▶In this study, we examined the inhibitory effect of sulforaphane on
tyrosinase
▶key enzymes in melanogensis
tyrosinase, tyrosinase-related protein 1 (TRP-1), tyrosinase-related protein 2 (TRP-2)
▶MAP kinase family
controls melanogenesis.
<induction> phosphorylated ERK
<inhibition> phosphorylated p38
MITF
melanin synthesis
▶skin is exposed to UV
1) αMSH
2) αMSH + MC1R(receptor)
3) MITF
4) tyrosinase expression

6.  Cell culture : B16 murine melanoma cells
 Measurement of melanin contents
 Cell viability assay : electron microscope
 Tyrosinase activity assay : Bradford assay, microplate reader
 Western blotting analysis
 Statistical analysis : Dunnett’s test
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7. Fig. 1. Effects of Sulforaphane on
Melanin Production in B16 Melanoma
Cells.
Effects of sulforaphane on melanogenesis using B16 melanoma cells
Fig. 2. Cytotoxic Effects of Sulforaphane
on B16 Melanoma Cells.
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“melanin 합성의 저해!!”
“no cytotoxic effects”

8. Fig. 3. Effects of Sulforaphane on
Tyrosinase Activity in B16 Melanoma Cells.
Effects of sulforaphane on tyrosinase expression in B16 melanoma cells
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▶in vitro tyrosinae assay
 tyrosinase의 활성에 영향을
주지 않았다.
▶cellular tyrosinase assay
 tyrosinase activity가 감소
tyrosinase의 수량이 감소!

9. Fig. 4. Effects of Sulforaphane on Tyrosinase Expression in B16 Melanoma Cells.
Effects of sulforaphane on tyrosinase expression in B16 melanoma cells
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Fig. 5. Effects of Sulforaphane on MAP Kinase Pathways in B16 Melanoma Cells.
Effects of sulforaphane on MAP kinase activation in B16 melanoma cells
sulforaphane
tyrosinase
p-p38 (360-720 min)
p-ERK (60-720 min)

10. 10
Sulforaphane treatment inhibited p38 phosphorylation in B16 cells.
1. inhibition of p38 phosphorylation
induction of ERK phosphorylation
2. inhibition of MITF activation
3. inhibition of TRY, TRP1, TRP2 expression
4. inhibition of melanin synthesis
Sulforaphane