인체 멜라닌 세포의 멜라닌형성에 대한 WIF-1의 효과
Wnt Inhibitory Factor (WIF)-1 Promotes Melanogenesis in Normal Human Melanocytes
- 주제(키워드) melanocyte , melanogenesis , WIF-1 , pigmentation , Wnt antagonist
- 발행기관 아주대학교
- 지도교수 강희영
- 발행년도 2011
- 학위수여년월 2011. 8
- 학위명 석사
- 학과 및 전공 일반대학원 의생명과학과
- 파일정보 워드
- 실제URI http://www.dcollection.net/handler/ajou/000000011924
- 본문언어 영어
- 저작권 아주대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
- ABSTRACT - Wnt Inhibitory Factor (WIF)-1 Promotes Melanogenesis in Human Melanocytes Background: Wnt inhibitory factor-1 (WIF-1) acts as a suppressor of Wnt antagonists. There is no study available on melanogenesis effect of human melanocytes. Objectives: In this study, we investigated the expression of WIF-1 in human melanocytes and the effects of WIF-1 on melanogenesis. Method: To determinutese whether WIF-1 is expressed in human melanocytes, we used immunocytochemistry, immunhistochemistry, western blot analysis and reverse transcription polymerase chain reaction. The cell proliferation was measured using the coulter counter. The effects on pigmentation were investigated with measurement of melanin contents, tyrosinase activity and its expression. Result: WIF-1 was expressed in melanocytes. WIF-1 inhibited proliferation of human melanocytes in a dose-dependent manner. WIF-1 also appeared to stimulate the melanin synthesis of human melanocytes. This increase in pigmentation was due to stimulation of the tyrosinase activity and increased expression of MITF and tyrosinase. Conclusion: We demonstrated that WIF-1 was expressed in human melanocytes. Our findings suggest that WIF-1 may play a role in melanogenesis. Key Words: melanocyte, melanogenesis, WIF-1, pigmentation, Wnt antagonist
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TABLE OF CONTENTS
ABSTRACT 5
TABLE OF CONTENTS 6
LIST OF FIGURES 8
LIST OF TABLES 9
I. INTRODUCTION 10
A. Human melanocyte development and pigmentati on 10
B. Wnt signaling pathway 12
C. The Wnt family 14
D. The Wnt antagonist 15
E. Melanocyte and Wnt signaling 18
II. MATERIALS AND METHODS 19
A. Human melanocyte culture 19
B. B16 melanoma cell lines culture 19
C. Human fibroblast culture 20
D. Human keratinocyte culture 20
E. Cell growth assay 20
F. Melanin content determinutesation 12
G. Tyrosinase activity assay 21
H. Skin organ culture 21
I. Pigmentation assay in cultured skin 21
J. Immunocytochemistry 22
K. Western blottiong 23
L. WIF-1 transfection 23
M. Reverse transcriptase-polymerase chain reaction (RT-PCR) 24
N. Statistical analysis 24
III. RESULTS 25
A. WIF-1 is expressed in human melanocytes, fibroblasts and keratinocytes 25
B. WIF-1 increases pigmentation of cultured human skin 27
C. WIF-1 treatment enhances pigmentation in B16 melanoma cells 29
D. WIF-1 treatment enhances pigmentation in human melanocytes 32
E. WIF-1 transfection induces pigmentation in B16 melanoma cells 35
F. WIF-1 transfection induces pigmentation in human melanocytes 38
IV. DISCUSSION 41
V. CONCLUSION 42
REFERENCES 43
국문요약 49
LIST OF FIGURES
Fig. 1. Melanogenesis 11
Fig. 2. Wnt signaling pathways 13
Fig. 3. WIF-1 is expressed in human melanocytes, fibroblasts and keratinocytes 26
Fig. 4. WIF-1 increases pigmentation of cultured human skin 28
Fig. 5. WIF-1 treatment enhances pigmentation in B16 melanoma cells 30
Fig. 6. WIF-1 treatment enhances pigmentation in human melanocytes 33
Fig. 7. WIF-1 transfection induces pigmentation in B16 melanoma cells 36
Fig. 8. WIF-1 transfection induces pigmentation in human melanocytes 39
LIST OF TABLES
Table 1. Wnt antagonist family molecules 17
Table 2. Effects of WIF-1 on proliferation, melanin contents and tyrosinase activity of B16 melanoma cells 31
Table 3. Effects of WIF-1 on proliferation, melanin contents and tyrosinase activity of human melanocytes 34
Table 4. Effects of WIF-1 transfection on proliferation, melanin contents and tyrosinase activity of B16 melanoma cells 37
Table 5 . Effects of WIF-1 transfection on proliferation, melanin contents and tyrosinase activity of human melanocytes 40
