InnVit基因对黑素细胞生物学功能的影响及其分子机制
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摘要
白癜风是一种色素脱失性皮肤病,以表皮、粘膜和毛发的黑素细胞脱失形成的白斑、灰白发为主要症状,其发病机制存在自身免疫学、氧化应激学、神经学以及遗传学等多种假说。近年来,国内外大量研究表明白癜风的发病与遗传因素密切相关,可能是一种多基因遗传病。
据报道InnVit基因是与白癜风患者的黑素细胞表达下调相关的新基因。通过序列比对发现InnVit基因为缺少81bp内含子的FBXO11基因。为了探讨InnVit基因在黑素细胞中的生物学功能,明确InnVit基因在白癜风中的作用,本论文构建了沉默InnVit基因的siRNA及InnVit基因过表达载体p3XF-P120,研究InnVit基因的抑制和过表达对黑素细胞生物学活性及酪氨酸酶输出的影响。
论文通过设计构建了人工合成的特异性siRNA,同时构建了该基因的过表达质粒,同时分别转染黑素细胞,从而研究InnVit基因抑制和过表达对黑素细胞生物学功能及黑素细胞酪氨酸酶表达及内质网输出的的影响。
首先,检测了InnVit基因mRNA和蛋白的表达,siRNA的抑制作用显著降低了InnVit基因mRNA和蛋白的表达量,p3XF-P120质粒的过表达显著增加了InnVit基因mRNA和蛋白的表达量。其次,InnVit基因抑制和过表达对黑素细胞增殖、凋亡及迁移相关生物学功能的影响。采用siRNA抑制和质粒载体过表达后,InnVit基因的过表达促进了细胞增殖,抑制细胞凋亡;InnVit基因的抑制使黑素细胞会出现从G0/G1期到S期的细胞周期停滞现象,极大地降低了细胞增殖和存活的能力,同时显著增加黑素细胞的早期凋亡水平。细胞迁移能力及酶活性检测表明,siRNA抑制和过表达InnVit基因对细胞迁移力及2金属蛋白酶MMP9和MMP的表达没有影响。推测,InnVit基因的抑制主要通过促进细胞凋亡和细胞周期停滞而降低黑素细胞的存活和增殖能力。
最后,研究了InnVit基因对黑素细胞内质网膨胀与酪氨酸酶的相关性。扫描电镜观察表明,siRNA抑制的黑素细胞内质网有明显的膨胀扩大现象,其酪氨酸酶蛋白表达存在极显著的增加。细胞免疫荧光共定位了酪氨酸酶和内质网标记钙网蛋白,发现抑制组细胞的酪氨酸酶大部分停留在内质网中,表明其酪氨酸酶的内质网输出发生阻滞,可见抑制InnVit基因表达影响了黑素细胞的酪氨酸酶从内质网输出,导致了细胞内质网膨胀,从而导致细胞生物学活性异常。
以上研究表明,本研究为InnVit基因影响黑素细胞的增殖和凋亡能力提供了有力的证据,并揭示了白癜风黑素细胞酪氨酸酶的内质网功能性输出障碍的可能机制。
Vitiligo is a common autoimmune, frequent family clustering depigmentary disorder of skin that results from destruction melanocytes. To date, the pathogenesis of vitiligo is unknown although there are many theories about its etiology, including autoimmune, neural and genetic hypotheses. In recent years, genetic contributions were revealed as the most important causative factor of vitiligo. Moreover, scientists holding the other hypotheses of vitoligo began to seek supporting evidence from genetics.
InnVit, a novel gene, was reportedly downregulated in vitiligo melanocytes. Sequence analysis revealed that the InnVit gene lacking its intron is the FBXO11 gene. To analyze the effect of InnVit gene on MC biological behavior further investigates molecular mechanisms of InnVit gene on Tyrosinase. We designed and synthesized siRNA and expression vectors for the InnVit gene, Here, we further investigate the effects of inhibition and over-expression of InnVit gene on the biological behavior of melanocytes, and on the export of Tyrosinase from ER.
First, transfection efficiency was estimated by RT-PCR and western blot analysis at post-transfection, Silencing of InnVit gene remarkly decreased the expression of mRNA and protein of InnVit,while over-expression of InnVit gene increased the expression of mRNA and protein of InnVit .
Second,We detected biological behavior of melanocytes on cell proliferation, apoptosis, transwell and melanin synthesis. The results demonstrated that the silencing of InnVit gene inhibited cell proliferation and induced cell apoptosis efficiently, while the over-expression of InnVit gene could promote cell proliferation and suppress cell apoptosis. Silencing of InnVit gene resulted in a decreased ability to progress from G0/G1 phase to S phase, eventually inhibiting cell proliferation, which was confirmed by MTT assay. Silencing or over-expression of InnVit gene has little effect on cell migration (as determined by transwell assay) or MMP9 and MMP2 activity.
Last, our study further identified the effects of InnVit gene on melanocytes and the relationship of between dilated ER and Tyrosinase by inhibition and over-expression of InnVit gene.By electron microscopicy, obvious swelling of the endoplasmic reticulum (ER) was found in the cells transfected by siRNA for InnVit. Protein levels of Tyrosinase were extraordinarily high following inhibition of InnVit. Further examination revealed Tyrosinase and Calreticulin were co-localized in ER of the transfected cells following siRNA of InnVit, indicating that Tyrosinase could not export from ER effectively, consequently in the abnormity of biological behavior of melanocytes.
Collectively, this study provides support that InnVit plays an important role in regulating the proliferation and apoptosis of melanocytes, and revealed the probably mechanisms of functional export of Tyrosinase from ER in vitiligo melanocytes.
据报道InnVit基因是与白癜风患者的黑素细胞表达下调相关的新基因。通过序列比对发现InnVit基因为缺少81bp内含子的FBXO11基因。为了探讨InnVit基因在黑素细胞中的生物学功能,明确InnVit基因在白癜风中的作用,本论文构建了沉默InnVit基因的siRNA及InnVit基因过表达载体p3XF-P120,研究InnVit基因的抑制和过表达对黑素细胞生物学活性及酪氨酸酶输出的影响。
论文通过设计构建了人工合成的特异性siRNA,同时构建了该基因的过表达质粒,同时分别转染黑素细胞,从而研究InnVit基因抑制和过表达对黑素细胞生物学功能及黑素细胞酪氨酸酶表达及内质网输出的的影响。
首先,检测了InnVit基因mRNA和蛋白的表达,siRNA的抑制作用显著降低了InnVit基因mRNA和蛋白的表达量,p3XF-P120质粒的过表达显著增加了InnVit基因mRNA和蛋白的表达量。其次,InnVit基因抑制和过表达对黑素细胞增殖、凋亡及迁移相关生物学功能的影响。采用siRNA抑制和质粒载体过表达后,InnVit基因的过表达促进了细胞增殖,抑制细胞凋亡;InnVit基因的抑制使黑素细胞会出现从G0/G1期到S期的细胞周期停滞现象,极大地降低了细胞增殖和存活的能力,同时显著增加黑素细胞的早期凋亡水平。细胞迁移能力及酶活性检测表明,siRNA抑制和过表达InnVit基因对细胞迁移力及2金属蛋白酶MMP9和MMP的表达没有影响。推测,InnVit基因的抑制主要通过促进细胞凋亡和细胞周期停滞而降低黑素细胞的存活和增殖能力。
最后,研究了InnVit基因对黑素细胞内质网膨胀与酪氨酸酶的相关性。扫描电镜观察表明,siRNA抑制的黑素细胞内质网有明显的膨胀扩大现象,其酪氨酸酶蛋白表达存在极显著的增加。细胞免疫荧光共定位了酪氨酸酶和内质网标记钙网蛋白,发现抑制组细胞的酪氨酸酶大部分停留在内质网中,表明其酪氨酸酶的内质网输出发生阻滞,可见抑制InnVit基因表达影响了黑素细胞的酪氨酸酶从内质网输出,导致了细胞内质网膨胀,从而导致细胞生物学活性异常。
以上研究表明,本研究为InnVit基因影响黑素细胞的增殖和凋亡能力提供了有力的证据,并揭示了白癜风黑素细胞酪氨酸酶的内质网功能性输出障碍的可能机制。
Vitiligo is a common autoimmune, frequent family clustering depigmentary disorder of skin that results from destruction melanocytes. To date, the pathogenesis of vitiligo is unknown although there are many theories about its etiology, including autoimmune, neural and genetic hypotheses. In recent years, genetic contributions were revealed as the most important causative factor of vitiligo. Moreover, scientists holding the other hypotheses of vitoligo began to seek supporting evidence from genetics.
InnVit, a novel gene, was reportedly downregulated in vitiligo melanocytes. Sequence analysis revealed that the InnVit gene lacking its intron is the FBXO11 gene. To analyze the effect of InnVit gene on MC biological behavior further investigates molecular mechanisms of InnVit gene on Tyrosinase. We designed and synthesized siRNA and expression vectors for the InnVit gene, Here, we further investigate the effects of inhibition and over-expression of InnVit gene on the biological behavior of melanocytes, and on the export of Tyrosinase from ER.
First, transfection efficiency was estimated by RT-PCR and western blot analysis at post-transfection, Silencing of InnVit gene remarkly decreased the expression of mRNA and protein of InnVit,while over-expression of InnVit gene increased the expression of mRNA and protein of InnVit .
Second,We detected biological behavior of melanocytes on cell proliferation, apoptosis, transwell and melanin synthesis. The results demonstrated that the silencing of InnVit gene inhibited cell proliferation and induced cell apoptosis efficiently, while the over-expression of InnVit gene could promote cell proliferation and suppress cell apoptosis. Silencing of InnVit gene resulted in a decreased ability to progress from G0/G1 phase to S phase, eventually inhibiting cell proliferation, which was confirmed by MTT assay. Silencing or over-expression of InnVit gene has little effect on cell migration (as determined by transwell assay) or MMP9 and MMP2 activity.
Last, our study further identified the effects of InnVit gene on melanocytes and the relationship of between dilated ER and Tyrosinase by inhibition and over-expression of InnVit gene.By electron microscopicy, obvious swelling of the endoplasmic reticulum (ER) was found in the cells transfected by siRNA for InnVit. Protein levels of Tyrosinase were extraordinarily high following inhibition of InnVit. Further examination revealed Tyrosinase and Calreticulin were co-localized in ER of the transfected cells following siRNA of InnVit, indicating that Tyrosinase could not export from ER effectively, consequently in the abnormity of biological behavior of melanocytes.
Collectively, this study provides support that InnVit plays an important role in regulating the proliferation and apoptosis of melanocytes, and revealed the probably mechanisms of functional export of Tyrosinase from ER in vitiligo melanocytes.
引文
1、Nath SK, Majumder PP, andNordlund JJ. Genetic epidemiology of vitiligo: multilocus recessivity cross-validated [J]. Am.J.Hum.Genet, 1994, 55: 981-990.
2、Shajil EM, Chatterjee S, Agrawal D, et al.Vitiligo: Pathomechanisms and genetic polymorphism of susceptibility genes [J]. Ind.J.Exp.Biol. 2006, 44: 526-53.
3、Majumder PP, Nordlund JJ, and Nath SK. Pattern of Familial aggregation of vitiligo [J]. Arch.Dermatol. 1993, 129: 994-998.
4、Sun X, Xu A.,Wei X, et al.Geneticepidemiology of vitiligo: a study of 815 pro-bands and theirf amilies from southChina[J]. Int.J.Dermatol, 2006, 45: 1176-1181.
5、Gramer SF. The origin of epidermal melanocytes, implication for the histogenesis of nevi and melanomas [J]. Arch Path Lab Med,1 991, 115∶1 15-119.
6、Jose N, Rodriguez L, Jose T, et al. Analysis of a kinetic model for melanin biosynthesis pathway [J]. J Biol Chem,1992, 267∶3801-3810.
7、Nishioka E, Funasaka Y, Kondoh H, et al. Expression of tyrosinase, TRP-1 and TRP-2 in ultraviolet-irradiated human melanomas and melanocytes: TRP-2 protects melanoma cells from ultraviolet B induced apoptosis [J]. Melanoma Res, 1999, 9(5): 433-443.
8、Boissy RE, Nordlund JJ. Molecular basis of congenital hypopigmentary disorders in humans [J]. Pigment Cell Res, 1997, 10(1-2): 12-24.
9、Jimbow K, Chen H, Park JS, et al. Increased sensitivity of melanocytes to oxidative stress and abnormal expression of tyrosinase-related protein in vitiligo[J]. Br J Dermatol, 2001 Jan, 144(1): 55-65.
10、Casp CB, She JX, McCormack WT. Genetic association of the catalase gene (CAT) with vitiligo susceptibility [J]. Pigment Cell Res, 2002, 15(1): 62-66.
11、Cucchi ML, Frattini P, Santagostino G., et al. Higher plasma catecholamine andmetabolite levels in the early phase of non-segmental vitiligo [J]. Pigment Cell Res, 2000, 13: 28–32.
12、Lerner, A.B., Vitiligo. J. Invest [J]. Dermatol. 1959, 32: 285–310.
13、Moretti S, Spallanzani A, Amato L, Hautmann G., et al. New insights into the pathogenesis of vitiligo: imbalance of epidermal cytokines at sites of lesions [J]. Pigment Cell Res, 2002, 15: 87–92.
14、Garbelli S, Mantovani S, Palermo B, et al. Melanocyte-specific, cytotoxic T cell responses in vitiligo: the effective variant of melanoma immunity [J]. Pigment Cell Res, 2005, 18: 234–242.
15、Ongenae K, Van Geel N, and Naeyaert JM. Evidence for an autoimmune pathogenesis of vitiligo [J]. Pigment Cell Res, 2003, 16: 90–100.
16、Wiete W, Marcod I. Vitiligo puzzle: the pieces fall in place [J]. PigmentCellRes, 20: 345-359.
17、Casp CB, Litherland SA, She J, et al.Antigen processing and presentation genes invitiligo susceptibility [J]. FASEB Journal, 2002, 16(5): 22,1219.
18、Sander CS, Chang H, Hamm F, et al. Role of oxidative stress and the antioxidant network in cutaneous carcinogenesis [J]. Int J Dermatol, 2004, 43: 326–335.
19、Schallreuter, K.U. Oxidative stress in the human epidermis.G. Ital. Dermatol [J]. Venereol, 2005, 140: 505–514.
20、Dell’Anna ML, and Picardo M. A review and a new hypothesis for non-immunological pathogenetic mechanisms in vitiligo [J]. Pigment Cell Res, 2006, 19: 406–411.
21、Gauthier Y, Cario Andre M., and Taieb A. A critical appraisal of vitiligo etiologic theories. Is melanocyte loss a melanocytorrhagy [J]. Pigment Cell Res, 2003, 16: 322–332.
22、Slominski, A, Paus,R, Bomirski, A.Hypothesis: a possible role of the melatonin receptor in vitiligo [J]. J Roy Soc Med, 1989, 82: 539–541.
23、Alkhateeb A, Fain PR, Thody A,et al.Epidemiology of vitiligo and associated autoimmune diseases in Caucasian probands and their families [J]. Pigment CellResearch, 2003, 16(3): 208-214.
24、Tripathi RK, Flanders DJ, Young TL, et al. Microphthalmia-associated transctiption factor (MITF) locus lacks linkage to human vitiligo or osteopetrosis: an evaluation [J]. Pigment Cell Res, 1999, 12(3): 187-192.
25、Smith SB, Zhou BK, Orlow SL. Expression of tyrosinase and the tyrosinase related proteins in the Mitfvit (vitiligo) mouse eye: implications for the function of the microphthalmia transcription factor [J]. Exp Eye Res, 1998, 66(4): 403-410.
26、Lamoreux ML, Boissy RE, Womack JE, et al. The vit gene maps to the mi (microphthalmia) locus of the laboratory mouse [J]. J Hered, 1992, 83(6): 435-439.
27、Richards KA, Fukai K, Oiso N, et al. A novel KIT mutation results in piebaldism with progressive depigmentation [J]. J Am Acad Dermatol, 2001, 44(2): 288-292.
28、Park HH, Ha E, UhmYK, et al. Association study between catalase gene polymorphisms and the susceptibility to vitiligo in Korean population [J]. Experimental Dermatology, 2006; 15(5): 377-380.
29、UhmYK, Yoon SH, Kang IJ, et al. Association of glutathione Stransferase gene polymorphisms (GSTM1 and GSTT1) of vitiligo in Korean population [J]. Life Sci, 2007, 81(3): 223-227.
30、Jin SY, Park HH, Li GZ, et al. Association of estrogen receptor 1intron 1 C/T polymorphism in Korean vitiligo patients [J]. J Dermatol Sci 2004, 35(3): 181-186.
31、卢宁,高天文.雌激素受体ER-α基因多态性与白癜风相关性研究[J].中国皮肤性病学杂志, 2007, 21: 147-149.
32、Uitter linden AG, Fang Y, VanMeurs JB, et al. Genetics and biology of vitamin D receptor polymorphisms [J]. Gene 2004, 338(2): 143-156.
33、Watabe H, Soma Y, Kawa Y, et al. Differentiation of murine melanocyte precursors induced by 1, 25-dihydroxy vitamin D3 is associated with the stimulation of endothelin B receptor expression [J]. J Invest Dermatol, 2002,119(3): 583-589.
34、Birlea S, Birlea M, Cimponeriu D, et al. Autoimmune diseasesand vitamin D receptor Apa-I polymorphism are associated with vitiligo in a small inbred Romanian community [J]. Acta DermVenereol 2006, 86(3): 209-214.
35、Li C, Liu Z, Zhang Z, et al. Genetic variants of the vitami D receptor gene alter risk of cutaneous melanoma [J]. J Invest Dermatol, 2007, 127(2): 276-280.
36、杨励,王雷,孙林潮,等.维生素D受体基因多态性与白癜风的相关性研究[J].中国美容医学2007, 16(3): 302-304.
37、Tursen U, Kaya TI, Derici ME, et al.Association between cate-chol-O-methyltransferase polymorphism and vitiligo [J]. Arch Dermatol Res, 2002, 294: 143-146.
38、李凯,高琳,杨励,等.COMT基因单核苷酸多态性与汉族白癜风发病的相关性[J].中国美容医学, 2007, 16: 365-368.
39、Robinson J, WallerMJ, Parham P, et al. IMGT/HLA Database-a sequence database for the human major histo compatibility complex [J]. Nucleic Acids Res, 2001, 29(1): 210-213.
40、Little AM, Parham P. Polymorphism and evolution of HLA class I and II genes and molecules [J].Rev Immunogenet, 1999, 1(1): 105-123.
41、Zamani M, SpaepenM, Sghar SS, et al. Linkage and association of HLA class II genes with vitiligo in a Dutch population [J]. Br J Dermatol 2001, 145(1): 90-94.
42、汪继云,杨森,王红艳,等.HLA-DQA1和HLA-DQBI等位基因与皖籍汉族人群白癜风的相关性[J].中华皮肤科杂志, 2005, 38(6): 357-359.
43、Alkhateeb A, Stetler GL, Old W,et al.Mapping of an autoimmunity susceptibility locus (AIS1) to chromosome 1p31.3-p32.2 [J].Human Molecular Genetics, 2002, 11(6): 661-667.
44、Zhang XJ, Chen JJ, Liu Jb. The genetic concept of vitiligo [J]. J Dermatol Sci, 2005, 39: 137-146.
45、Fain PR,Gowan K, LaBerge GS,et al.A genomewide screen for generalized vitiligo:confirmation of AIS1 on chromosome 1p31 and evidence for additionalsusceptibility loci [J]. American Journal of Human Genetics, 2003, 72(6): 1560-1564.
46、Na GY, Lee KH, Kim MK,et al.Polymorphisms in the melanocortin-1 receptor( MC1R ) and agouti signaling protein(ASIP) genes in Korean vitiligo patients [J]. Pigment Cell Research, 2003, 16(4): 383-387.
47、Bradley CA, Kristensen D, She J,et al.Genetic association studies of immune response genes and vitiligo susceptibility [J].FASEB Journal, 2001, 15(4):7, 323.
48、Fain PR, Gowan K, LaBerge GS, et al. A genomewide Screen for generalized vitiligo: confirmation of AIS1 on chromo-some1p31 and evidence for additional susceptibility loci [J]. Am. J Hum Genet, 2003, 72, 1560–1564.
49、Jin Y, Mailloux CM, Gowan K, et al. NALP1 in vitiligo-associated multiple autoimmune disease [J]. N Engl J Med, 2007, 356: 1216–1225.
50、Jin Y, Birlea SA, Fain PR, et al. Genetic Variations in NALP1 Are Associated with Generalized Vitiligo in a Romanian Population [J]. J Invest Dermatol, 2007, 19, 127(11): 2558-62.
51、Jin Y, Riccardi SL, Gowan K, Fain PR, Spritz RA. Fine-Mapping of Vitiligo Susceptibility Loci on Chromosomes 7 and 9 and Interactions with NLRP1 (NALP1) [J]. J Invest Dermatol. 2009 Sep 3, Epub.
52、Kummer JA, Broekhuizen R, Everett H, et al. Inflammasome components NALP1 and 3 show distinct but separate expression profiles in human tissues suggesting as ite-specific role in the inflammatory response [J]. J Histochem Cytochem, 2007, 55: 443– 452.
53、Birlea SA, Gowan K, Fain PR, Spritz RA.Genome-Wide Association Study of Generalized Vitiligo in an Isolated European Founder Population Identifies SMOC2, in Close Proximity to IDDM8.J Invest Dermatol [J]. 2009 Nov 5, Epub.
54、Le Poole IC, Sarangarajan R, Zhao Y, et al.“VIT1”, a novel gene associated with vitiligo [J]. Pigment Cell Res, 2001, 14(6): 475-484.
55、Li YW, Xu AE, Wei XD, et al. Analysis of molecular character of gene“VIT1”in Chinese patients with vitiligo(abstr) [J]. J Invest Dermatol, 2006, 126(4):1-156.
56、Jin, J. Cardozo, T, Lovering, RC, et al. Systematic analysis and nomenclature of mammalian F-box proteins [J]. Genes Dev, 2004, 18: 2573-2580.
57、Jin J, Ang XL, Shirogane T, et al. Identification of substrates for f-box proteins [J]. Methods Enzymol, 2005, 399: 287-309.
58、Wassim M, Anatoly N. FBXO11 Promotes the Neddylation of p53 and Inhibits Its Transcriptional Activity [J]. J Biological Chem, 2007, 282: 1197-1804.
59、李永伟,张迪敏,许爱娥,等.白癜风相关基因-1的确认及其分子结构分析[J].中华皮肤科杂志, 2006, 39: 190-192.
60、Tobin DJ, Swanson NN, Pittelkow MR, et al. Melanocytes are not absent in lesional skin of long duration vitiligo [J]. J Pathol, 2000, 191: 407-416.
61、Friedlander R, Jarosch E, Urban J, et al. A regulatory link between ER-associated protein degradation and unfolded-protein response [J]. Nat cell boil, 2000, 2: 379-384.
62、Hiller MM, Finger A, Schweiger M, et al. ER degradation of a misfolded lumenal protein by the cytosolic ubiquitin-proteasome pathway [J]. Science, 1996, 273: 1725-1728.
63、Hershko A. Ciechanover, Varshavsky A. The ubiquitin system [J]. Nature Med, 2000, 6: 1073-1081.
64、Hershko A. The ubiquitin system for protein degradation and some of its roles in the control of the cell division cycle [J]. Cell Death Differ, 2005, 12: 1191-1197.
65、Orlowski RZ.The role of the ubiquitin-proteasome pathway in apoptosis [J]. Cell Death Differ, 1999, 6: 303-313.
66、Cope GA, Deshaies RJ. Targeted silencing of Jab1/Csn5 in human cells downregulates SCF activity through reduction of F-box protein levels [J]. BMC Biochem, 2006, 9: 123-134.
67、Minella AC, Clurman BE. Mechanisms of tumor suppression by the SCF (Fbw7) [J]. Cell Cycle, 2005, 4: 1356-1359.
68、Ruth H, Elaine C, Sherri S, et al. Proper folding and endoplasmic reticulum to golgi transport of tyrosinase are induced by its substrates, DOPA and tyrosine [J]. J Biological Chem, 2001, 276: 11933-11938.
69、Hu DN, Steven AM, Seth JO, Susana R, et al. Melanogenesis by human uvealmelanocytes in vitro [J]. Inveast Ophthalmology and Visual Sci, 1995, 36 (5): 931-938.
70、Hu DN. Methodology for Evaluation of Melanin Content and Production of Pigment Cells in Vitro [J]. Photochemistry and Photobiology, 2007, 83: 1–5.
71、Zhang XJ, Chen JJ, Liu JB. The genetic concept of vitiligo [J]. J Dermatol Sci, 2005, 39(3): 137-146.
72、李永伟,张迪敏,刘东银,等. InnVit基因沉默对黑素细胞B10BR黑素合成和细胞凋亡的影响[J].中国皮肤性病学杂志, 2007, 21(1):3-4.
73、Li YW, Chen FF, Lin FQ, et al.VIT1/FBXO11 knockdown induces morphological alterations and apoptosis in B10BR mouse melanocytes [J]. Int J Mol Med, 2009, 23(5):673-8.
74、Boissy RE, Liu YY, Medrano EE, et al. Structural aberration of the rough endoplasmic reticulum and melanosome compartmentalization in long-term cultures of melanocytes from vitiligo patients [J]. J Invest Dermatol, 1991, 97: 395-404.
75、Boissy RE, Beato KE, Nordlund JJ. Dilated rough endoplasmic reticulum and premature death in melanocytes cultured from the vitiligo mouse [J]. Am J Pathol, 1991, 138: 1511-1525.
76、Abida WM., Nikolaev A, Zhao WH, et al. FBXO11 promotes the neddylation of p53 and inhibits its transcriptional activity [J]. J biological chemistry, 2006, 282: 1797-1804.
77、King R W, Deshaies R J, Peters J M,et a1. Howproteolysis drives the cell cycle [J]. Science, 1996, 274:1652-1659.
78、Peters J M. SCF and APC: the Yin and Yang of cell cycle regulated proteolysis [J]. Curr Opin Cell Biol, 1998, 10:759-768.
79、孙秀坤,许爱娥.黑素细胞内酪氨酸酶翻译后加工运输某些色素病的关系[J].国外医学皮肤病学分册, 2005, 31: 50-52.
80、Le Poole IC, Boissy RE., Sarangarajan R, et al. PIG3V, an immortalized human vitiligo melanocyte cell line, expresses dilated endoplasmic reticulum [J]. InVitro Cell Dev Bio Anim, 2000, 36:309-319.
2、Shajil EM, Chatterjee S, Agrawal D, et al.Vitiligo: Pathomechanisms and genetic polymorphism of susceptibility genes [J]. Ind.J.Exp.Biol. 2006, 44: 526-53.
3、Majumder PP, Nordlund JJ, and Nath SK. Pattern of Familial aggregation of vitiligo [J]. Arch.Dermatol. 1993, 129: 994-998.
4、Sun X, Xu A.,Wei X, et al.Geneticepidemiology of vitiligo: a study of 815 pro-bands and theirf amilies from southChina[J]. Int.J.Dermatol, 2006, 45: 1176-1181.
5、Gramer SF. The origin of epidermal melanocytes, implication for the histogenesis of nevi and melanomas [J]. Arch Path Lab Med,1 991, 115∶1 15-119.
6、Jose N, Rodriguez L, Jose T, et al. Analysis of a kinetic model for melanin biosynthesis pathway [J]. J Biol Chem,1992, 267∶3801-3810.
7、Nishioka E, Funasaka Y, Kondoh H, et al. Expression of tyrosinase, TRP-1 and TRP-2 in ultraviolet-irradiated human melanomas and melanocytes: TRP-2 protects melanoma cells from ultraviolet B induced apoptosis [J]. Melanoma Res, 1999, 9(5): 433-443.
8、Boissy RE, Nordlund JJ. Molecular basis of congenital hypopigmentary disorders in humans [J]. Pigment Cell Res, 1997, 10(1-2): 12-24.
9、Jimbow K, Chen H, Park JS, et al. Increased sensitivity of melanocytes to oxidative stress and abnormal expression of tyrosinase-related protein in vitiligo[J]. Br J Dermatol, 2001 Jan, 144(1): 55-65.
10、Casp CB, She JX, McCormack WT. Genetic association of the catalase gene (CAT) with vitiligo susceptibility [J]. Pigment Cell Res, 2002, 15(1): 62-66.
11、Cucchi ML, Frattini P, Santagostino G., et al. Higher plasma catecholamine andmetabolite levels in the early phase of non-segmental vitiligo [J]. Pigment Cell Res, 2000, 13: 28–32.
12、Lerner, A.B., Vitiligo. J. Invest [J]. Dermatol. 1959, 32: 285–310.
13、Moretti S, Spallanzani A, Amato L, Hautmann G., et al. New insights into the pathogenesis of vitiligo: imbalance of epidermal cytokines at sites of lesions [J]. Pigment Cell Res, 2002, 15: 87–92.
14、Garbelli S, Mantovani S, Palermo B, et al. Melanocyte-specific, cytotoxic T cell responses in vitiligo: the effective variant of melanoma immunity [J]. Pigment Cell Res, 2005, 18: 234–242.
15、Ongenae K, Van Geel N, and Naeyaert JM. Evidence for an autoimmune pathogenesis of vitiligo [J]. Pigment Cell Res, 2003, 16: 90–100.
16、Wiete W, Marcod I. Vitiligo puzzle: the pieces fall in place [J]. PigmentCellRes, 20: 345-359.
17、Casp CB, Litherland SA, She J, et al.Antigen processing and presentation genes invitiligo susceptibility [J]. FASEB Journal, 2002, 16(5): 22,1219.
18、Sander CS, Chang H, Hamm F, et al. Role of oxidative stress and the antioxidant network in cutaneous carcinogenesis [J]. Int J Dermatol, 2004, 43: 326–335.
19、Schallreuter, K.U. Oxidative stress in the human epidermis.G. Ital. Dermatol [J]. Venereol, 2005, 140: 505–514.
20、Dell’Anna ML, and Picardo M. A review and a new hypothesis for non-immunological pathogenetic mechanisms in vitiligo [J]. Pigment Cell Res, 2006, 19: 406–411.
21、Gauthier Y, Cario Andre M., and Taieb A. A critical appraisal of vitiligo etiologic theories. Is melanocyte loss a melanocytorrhagy [J]. Pigment Cell Res, 2003, 16: 322–332.
22、Slominski, A, Paus,R, Bomirski, A.Hypothesis: a possible role of the melatonin receptor in vitiligo [J]. J Roy Soc Med, 1989, 82: 539–541.
23、Alkhateeb A, Fain PR, Thody A,et al.Epidemiology of vitiligo and associated autoimmune diseases in Caucasian probands and their families [J]. Pigment CellResearch, 2003, 16(3): 208-214.
24、Tripathi RK, Flanders DJ, Young TL, et al. Microphthalmia-associated transctiption factor (MITF) locus lacks linkage to human vitiligo or osteopetrosis: an evaluation [J]. Pigment Cell Res, 1999, 12(3): 187-192.
25、Smith SB, Zhou BK, Orlow SL. Expression of tyrosinase and the tyrosinase related proteins in the Mitfvit (vitiligo) mouse eye: implications for the function of the microphthalmia transcription factor [J]. Exp Eye Res, 1998, 66(4): 403-410.
26、Lamoreux ML, Boissy RE, Womack JE, et al. The vit gene maps to the mi (microphthalmia) locus of the laboratory mouse [J]. J Hered, 1992, 83(6): 435-439.
27、Richards KA, Fukai K, Oiso N, et al. A novel KIT mutation results in piebaldism with progressive depigmentation [J]. J Am Acad Dermatol, 2001, 44(2): 288-292.
28、Park HH, Ha E, UhmYK, et al. Association study between catalase gene polymorphisms and the susceptibility to vitiligo in Korean population [J]. Experimental Dermatology, 2006; 15(5): 377-380.
29、UhmYK, Yoon SH, Kang IJ, et al. Association of glutathione Stransferase gene polymorphisms (GSTM1 and GSTT1) of vitiligo in Korean population [J]. Life Sci, 2007, 81(3): 223-227.
30、Jin SY, Park HH, Li GZ, et al. Association of estrogen receptor 1intron 1 C/T polymorphism in Korean vitiligo patients [J]. J Dermatol Sci 2004, 35(3): 181-186.
31、卢宁,高天文.雌激素受体ER-α基因多态性与白癜风相关性研究[J].中国皮肤性病学杂志, 2007, 21: 147-149.
32、Uitter linden AG, Fang Y, VanMeurs JB, et al. Genetics and biology of vitamin D receptor polymorphisms [J]. Gene 2004, 338(2): 143-156.
33、Watabe H, Soma Y, Kawa Y, et al. Differentiation of murine melanocyte precursors induced by 1, 25-dihydroxy vitamin D3 is associated with the stimulation of endothelin B receptor expression [J]. J Invest Dermatol, 2002,119(3): 583-589.
34、Birlea S, Birlea M, Cimponeriu D, et al. Autoimmune diseasesand vitamin D receptor Apa-I polymorphism are associated with vitiligo in a small inbred Romanian community [J]. Acta DermVenereol 2006, 86(3): 209-214.
35、Li C, Liu Z, Zhang Z, et al. Genetic variants of the vitami D receptor gene alter risk of cutaneous melanoma [J]. J Invest Dermatol, 2007, 127(2): 276-280.
36、杨励,王雷,孙林潮,等.维生素D受体基因多态性与白癜风的相关性研究[J].中国美容医学2007, 16(3): 302-304.
37、Tursen U, Kaya TI, Derici ME, et al.Association between cate-chol-O-methyltransferase polymorphism and vitiligo [J]. Arch Dermatol Res, 2002, 294: 143-146.
38、李凯,高琳,杨励,等.COMT基因单核苷酸多态性与汉族白癜风发病的相关性[J].中国美容医学, 2007, 16: 365-368.
39、Robinson J, WallerMJ, Parham P, et al. IMGT/HLA Database-a sequence database for the human major histo compatibility complex [J]. Nucleic Acids Res, 2001, 29(1): 210-213.
40、Little AM, Parham P. Polymorphism and evolution of HLA class I and II genes and molecules [J].Rev Immunogenet, 1999, 1(1): 105-123.
41、Zamani M, SpaepenM, Sghar SS, et al. Linkage and association of HLA class II genes with vitiligo in a Dutch population [J]. Br J Dermatol 2001, 145(1): 90-94.
42、汪继云,杨森,王红艳,等.HLA-DQA1和HLA-DQBI等位基因与皖籍汉族人群白癜风的相关性[J].中华皮肤科杂志, 2005, 38(6): 357-359.
43、Alkhateeb A, Stetler GL, Old W,et al.Mapping of an autoimmunity susceptibility locus (AIS1) to chromosome 1p31.3-p32.2 [J].Human Molecular Genetics, 2002, 11(6): 661-667.
44、Zhang XJ, Chen JJ, Liu Jb. The genetic concept of vitiligo [J]. J Dermatol Sci, 2005, 39: 137-146.
45、Fain PR,Gowan K, LaBerge GS,et al.A genomewide screen for generalized vitiligo:confirmation of AIS1 on chromosome 1p31 and evidence for additionalsusceptibility loci [J]. American Journal of Human Genetics, 2003, 72(6): 1560-1564.
46、Na GY, Lee KH, Kim MK,et al.Polymorphisms in the melanocortin-1 receptor( MC1R ) and agouti signaling protein(ASIP) genes in Korean vitiligo patients [J]. Pigment Cell Research, 2003, 16(4): 383-387.
47、Bradley CA, Kristensen D, She J,et al.Genetic association studies of immune response genes and vitiligo susceptibility [J].FASEB Journal, 2001, 15(4):7, 323.
48、Fain PR, Gowan K, LaBerge GS, et al. A genomewide Screen for generalized vitiligo: confirmation of AIS1 on chromo-some1p31 and evidence for additional susceptibility loci [J]. Am. J Hum Genet, 2003, 72, 1560–1564.
49、Jin Y, Mailloux CM, Gowan K, et al. NALP1 in vitiligo-associated multiple autoimmune disease [J]. N Engl J Med, 2007, 356: 1216–1225.
50、Jin Y, Birlea SA, Fain PR, et al. Genetic Variations in NALP1 Are Associated with Generalized Vitiligo in a Romanian Population [J]. J Invest Dermatol, 2007, 19, 127(11): 2558-62.
51、Jin Y, Riccardi SL, Gowan K, Fain PR, Spritz RA. Fine-Mapping of Vitiligo Susceptibility Loci on Chromosomes 7 and 9 and Interactions with NLRP1 (NALP1) [J]. J Invest Dermatol. 2009 Sep 3, Epub.
52、Kummer JA, Broekhuizen R, Everett H, et al. Inflammasome components NALP1 and 3 show distinct but separate expression profiles in human tissues suggesting as ite-specific role in the inflammatory response [J]. J Histochem Cytochem, 2007, 55: 443– 452.
53、Birlea SA, Gowan K, Fain PR, Spritz RA.Genome-Wide Association Study of Generalized Vitiligo in an Isolated European Founder Population Identifies SMOC2, in Close Proximity to IDDM8.J Invest Dermatol [J]. 2009 Nov 5, Epub.
54、Le Poole IC, Sarangarajan R, Zhao Y, et al.“VIT1”, a novel gene associated with vitiligo [J]. Pigment Cell Res, 2001, 14(6): 475-484.
55、Li YW, Xu AE, Wei XD, et al. Analysis of molecular character of gene“VIT1”in Chinese patients with vitiligo(abstr) [J]. J Invest Dermatol, 2006, 126(4):1-156.
56、Jin, J. Cardozo, T, Lovering, RC, et al. Systematic analysis and nomenclature of mammalian F-box proteins [J]. Genes Dev, 2004, 18: 2573-2580.
57、Jin J, Ang XL, Shirogane T, et al. Identification of substrates for f-box proteins [J]. Methods Enzymol, 2005, 399: 287-309.
58、Wassim M, Anatoly N. FBXO11 Promotes the Neddylation of p53 and Inhibits Its Transcriptional Activity [J]. J Biological Chem, 2007, 282: 1197-1804.
59、李永伟,张迪敏,许爱娥,等.白癜风相关基因-1的确认及其分子结构分析[J].中华皮肤科杂志, 2006, 39: 190-192.
60、Tobin DJ, Swanson NN, Pittelkow MR, et al. Melanocytes are not absent in lesional skin of long duration vitiligo [J]. J Pathol, 2000, 191: 407-416.
61、Friedlander R, Jarosch E, Urban J, et al. A regulatory link between ER-associated protein degradation and unfolded-protein response [J]. Nat cell boil, 2000, 2: 379-384.
62、Hiller MM, Finger A, Schweiger M, et al. ER degradation of a misfolded lumenal protein by the cytosolic ubiquitin-proteasome pathway [J]. Science, 1996, 273: 1725-1728.
63、Hershko A. Ciechanover, Varshavsky A. The ubiquitin system [J]. Nature Med, 2000, 6: 1073-1081.
64、Hershko A. The ubiquitin system for protein degradation and some of its roles in the control of the cell division cycle [J]. Cell Death Differ, 2005, 12: 1191-1197.
65、Orlowski RZ.The role of the ubiquitin-proteasome pathway in apoptosis [J]. Cell Death Differ, 1999, 6: 303-313.
66、Cope GA, Deshaies RJ. Targeted silencing of Jab1/Csn5 in human cells downregulates SCF activity through reduction of F-box protein levels [J]. BMC Biochem, 2006, 9: 123-134.
67、Minella AC, Clurman BE. Mechanisms of tumor suppression by the SCF (Fbw7) [J]. Cell Cycle, 2005, 4: 1356-1359.
68、Ruth H, Elaine C, Sherri S, et al. Proper folding and endoplasmic reticulum to golgi transport of tyrosinase are induced by its substrates, DOPA and tyrosine [J]. J Biological Chem, 2001, 276: 11933-11938.
69、Hu DN, Steven AM, Seth JO, Susana R, et al. Melanogenesis by human uvealmelanocytes in vitro [J]. Inveast Ophthalmology and Visual Sci, 1995, 36 (5): 931-938.
70、Hu DN. Methodology for Evaluation of Melanin Content and Production of Pigment Cells in Vitro [J]. Photochemistry and Photobiology, 2007, 83: 1–5.
71、Zhang XJ, Chen JJ, Liu JB. The genetic concept of vitiligo [J]. J Dermatol Sci, 2005, 39(3): 137-146.
72、李永伟,张迪敏,刘东银,等. InnVit基因沉默对黑素细胞B10BR黑素合成和细胞凋亡的影响[J].中国皮肤性病学杂志, 2007, 21(1):3-4.
73、Li YW, Chen FF, Lin FQ, et al.VIT1/FBXO11 knockdown induces morphological alterations and apoptosis in B10BR mouse melanocytes [J]. Int J Mol Med, 2009, 23(5):673-8.
74、Boissy RE, Liu YY, Medrano EE, et al. Structural aberration of the rough endoplasmic reticulum and melanosome compartmentalization in long-term cultures of melanocytes from vitiligo patients [J]. J Invest Dermatol, 1991, 97: 395-404.
75、Boissy RE, Beato KE, Nordlund JJ. Dilated rough endoplasmic reticulum and premature death in melanocytes cultured from the vitiligo mouse [J]. Am J Pathol, 1991, 138: 1511-1525.
76、Abida WM., Nikolaev A, Zhao WH, et al. FBXO11 promotes the neddylation of p53 and inhibits its transcriptional activity [J]. J biological chemistry, 2006, 282: 1797-1804.
77、King R W, Deshaies R J, Peters J M,et a1. Howproteolysis drives the cell cycle [J]. Science, 1996, 274:1652-1659.
78、Peters J M. SCF and APC: the Yin and Yang of cell cycle regulated proteolysis [J]. Curr Opin Cell Biol, 1998, 10:759-768.
79、孙秀坤,许爱娥.黑素细胞内酪氨酸酶翻译后加工运输某些色素病的关系[J].国外医学皮肤病学分册, 2005, 31: 50-52.
80、Le Poole IC, Boissy RE., Sarangarajan R, et al. PIG3V, an immortalized human vitiligo melanocyte cell line, expresses dilated endoplasmic reticulum [J]. InVitro Cell Dev Bio Anim, 2000, 36:309-319.