p16Ink4a in Melanocyte Senescence and Differentiation

Sviderskaya, Elena V.; Hill, Simon P.; Evans-Whipp, Tracy J.; Chin, Lynda; Orlow, Seth J.; Easty, David J.; Cheong, Sok Ching; Beach, David; DePinho, Ronald A.; Bennett, Dorothy C.

doi:10.1093/jnci/94.6.446

Cited by 166 publications

(163 citation statements)

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“…4A,B). As an indicator of cellular senescence, acidic senescence-associated ␤-galactosidase was mesured in wild-type and bcat sta melanocytes (Sviderskaya et al 2002). The number of senescence-associated ␤-galactosidase is clearly lower among bcat sta melanocytes (Fig.…”

Section: Bcat Sta Induces Immortalization In Culturementioning

confidence: 90%

“…As inactivation of the p16 INK4a gene is the most frequent cause of melanocyte immortalization (Sviderskaya et al 2002), we used immunofluorescence microscopy to study the effect of bcat sta on p16 INK4a protein abundance, as well as on the melanocyte differentiation markers melanin and Mitf, in primary melanocyte cultures 2, 6, and 20 wk after skin explantation (Fig. 5A,B).…”

Section: Bcat Sta Rapidly Represses P16 Ink4a Expression In Culturementioning

confidence: 99%

“…Western blots were performed as described previously (Sviderskaya et al 2002). The primary antibodies used were rabbit polyclonal anti-p16 INK4a antibody from Santa Cruz Biotechnology (SC-1207), mouse monoclonal anti-␤-catenin antibody from Transduction Laboratories (#610154), and mouse monoclonal anti-actin antibody from Euromedex (MAB1501).…”

Section: Western Blot Analysis and Immunofluorescence Microscopymentioning

confidence: 99%

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β-Catenin induces immortalization of melanocytes by suppressing p16^INK4a expression and cooperates with N-Ras in melanoma development

Delmas¹,

Beermann²,

Martinozzi³

et al. 2007

Genes Dev.

294

281

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Tumor progression is a multistep process in which proproliferation mutations must be accompanied by suppression of senescence. In melanoma, proproliferative signals are provided by activating mutations in NRAS and BRAF, whereas senescence is bypassed by inactivation of the p16 Ink4a gene. Melanomas also frequently exhibit constitutive activation of the Wnt/␤-catenin pathway that is presumed to induce proliferation, as it does in carcinomas. We show here that, contrary to expectations, stabilized ␤-catenin reduces the number of melanoblasts in vivo and immortalizes primary skin melanocytes by silencing the p16 Ink4a promoter. Significantly, in a novel mouse model for melanoma, stabilized ␤-catenin bypasses the requirement for p16 Ink4a mutations and, together with an activated N-Ras oncogene, leads to melanoma with high penetrance and short latency. The results reveal that synergy between the Wnt and mitogen-activated protein (MAP) kinase pathways may represent an important mechanism underpinning the genesis of melanoma, a highly aggressive and increasingly common disease.[Keywords: Mitf; Wnt; senescence; development; tumor suppressor; oncogene] Supplemental material is available at http://www.genesdev.org.

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Section: Bcat Sta Induces Immortalization In Culturementioning

confidence: 90%

Section: Bcat Sta Rapidly Represses P16 Ink4a Expression In Culturementioning

confidence: 99%

Section: Western Blot Analysis and Immunofluorescence Microscopymentioning

confidence: 99%

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β-Catenin induces immortalization of melanocytes by suppressing p16^INK4a expression and cooperates with N-Ras in melanoma development

Delmas¹,

Beermann²,

Martinozzi³

et al. 2007

Genes Dev.

294

281

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“…Immortal melanocyte cell lines melan-mu1, -2, and -3 and melan-pa1, -2, and -3 were grown from skins of neonatal muted (Muted mu/mu ) Ink4a-ArfϪ/Ϫ B6.CHMU/Le and pallid Ink4a-ArfϪ/Ϫ mice, respectively, as described previously (Sviderskaya et al, 2002). Melan-mu3 (referred to as melan-mu), melan-rp2 (Gwynn et al, 2004) (referred to as melan-rp; Bloc1s3 rp/rp ), melanpa1 (referred to as melan-pa), melan-coa2 (Suzuki et al, 2001) (referred to as melan-coa; Hps3 coa/coa ), and melan-a (Bennett et al, 1987) were maintained as described previously (Sviderskaya et al, 2002). Retrovirus production from transiently transfected 293T cells and transduction of primary melanocytes and cell lines were carried out as described previously (Swift et al, 1999).…”

Section: Cell Culture and Transgene Expressionmentioning

confidence: 99%

“…By contrast, nearly no pigment was observed in primary melanocytes from pallid mice, which lack the BLOC-1 subunit pallidin (Figure 1b), or in immortal melanocytes generated from muted (melan-mu; Figure 1d), pallid (melan-pa; see Figure 9c), or rp (melan-rp; our unpublished data) mice, which lack the BLOC-1 subunits muted, pallidin, and BLOS3, respectively. Modest pigmentation of primary pallid melanocytes developed only with increasing passage, likely due to senescence (Sviderskaya et al, 2002). On infection of early passage pallid melanocytes with recombinant retroviruses expressing hemagglutinin (HA)-epitope-tagged pallidin (PaHA), but not HA-tagged muted, ϳ10 -15% of the cells developed pigment within 3-5 d (Figure 1c).…”

mentioning

confidence: 99%

BLOC-1 Is Required for Cargo-specific Sorting from Vacuolar Early Endosomes toward Lysosome-related Organelles

Setty

Tenza

Truschel

et al. 2007

MBoC

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342

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Hermansky-Pudlak syndrome (HPS) is a genetic disorder characterized by defects in the formation and function of lysosome-related organelles such as melanosomes. HPS in humans or mice is caused by mutations in any of 15 genes, five of which encode subunits of biogenesis of lysosome-related organelles complex (BLOC)-1, a protein complex with no known function.Here, we show that BLOC-1 functions in selective cargo exit from early endosomes toward melanosomes. BLOC-1-deficient melanocytes accumulate the melanosomal protein tyrosinase-related protein-1 (Tyrp1), but not other melanosomal proteins, in endosomal vacuoles and the cell surface due to failed biosynthetic transit from early endosomes to melanosomes and consequent increased endocytic flux. The defects are corrected by restoration of the missing BLOC-1 subunit. Melanocytes from HPS model mice lacking a different protein complex, BLOC-2, accumulate Tyrp1 in distinct downstream endosomal intermediates, suggesting that BLOC-1 and BLOC-2 act sequentially in the same pathway. By contrast, intracellular Tyrp1 is correctly targeted to melanosomes in melanocytes lacking another HPS-associated protein complex, adaptor protein (AP)-3. The results indicate that melanosome maturation requires at least two cargo transport pathways directly from early endosomes to melanosomes, one pathway mediated by AP-3 and one pathway mediated by BLOC-1 and BLOC-2, that are deficient in several forms of HPS. INTRODUCTIONHermansky-Pudlak syndrome (HPS) is a genetic disorder characterized by hypopigmentation, prolonged bleeding, and sometimes ceroid accumulation, lung fibrosis, and/or immune defects leading to premature death Wei, 2006). HPS or a similar disorder in mice results from mutations in any of at least 15 genes (Wei, 2006). All of these genes are ubiquitously expressed, but their mutation in HPS affects mainly the generation and function of selected tissuespecific lysosome-related organelles (LROs;Bonifacino, 2004;Di Pietro and Dell'Angelica, 2005). Those LROs that are most severely affected in all forms of HPS-pigment cell melanosomes, platelet dense granules, and lung lamellar bodies-are unique in that they coexist with bona fide lysosomes in their respective cell types (Dell'Angelica et al., 2000;Marks and Seabra, 2001). The 15 known HPS-associated genes have been identified, and although the products of most are thought to participate in trafficking events that are uniquely required to form this class of LRO, the function of only a few is understood in detail.The genes disrupted in human HPS-7 (Li et al., 2003) and HPS-8 (Morgan et al., 2006) and in the mouse HPS models pallid, muted, reduced pigmentation (rp), cappuccino, and sandy encode five of the eight known subunits of a stable protein complex known as biogenesis of lysosome-related organelles complex (BLOC)-1 (Falcon-Perez et al., 2002;Moriyama and Bonifacino, 2002;Ciciotte et al., 2003;Li et al., 2003;Gwynn et al., 2004;Starcevic and Dell'Angelica, 2004). To date, no specific subcellular function has been assigned...

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Isolation, Culture, and Transfection of Melanocytes

et al. 2014

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Located in the basal epidermis and hair follicles, melanocytes of the integument are responsible for its coloration through production of melanin pigments. Melanin is produced in lysosomal‐like organelles called melanosomes. In humans, this skin pigmentation acts as an ultraviolet radiation filter. Abnormalities in the division of melanocytes are quite common, with potentially oncogenic growth usually followed by cell senescence producing benign naevi (moles), or occasionally melanoma. Therefore, melanocytes are a useful model for studying melanoma, as well as pigmentation and organelle transport and the diseases affecting these mechanisms. This chapter focuses on the isolation, culture, and transfection of human and murine melanocytes. The first basic protocol describes the primary culture of melanocytes from human skin and the maintenance of growing cultures. The second basic protocol details the subculture and preparation of mouse keratinocyte feeder cells. The primary culture of melanocytes from mouse skin is described in the third basic protocol, and, lastly, the fourth basic protocol outlines a technique for transfecting melanocytes and melanoma cells. Curr. Protoc. Cell Biol. 63:1.8.1‐1.8.20. © 2014 by John Wiley & Sons, Inc.

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p16Ink4a in Melanocyte Senescence and Differentiation

Cited by 166 publications

References 46 publications

β-Catenin induces immortalization of melanocytes by suppressing p16^INK4a expression and cooperates with N-Ras in melanoma development

β-Catenin induces immortalization of melanocytes by suppressing p16^INK4a expression and cooperates with N-Ras in melanoma development

BLOC-1 Is Required for Cargo-specific Sorting from Vacuolar Early Endosomes toward Lysosome-related Organelles

Isolation, Culture, and Transfection of Melanocytes

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p16Ink4a in Melanocyte Senescence and Differentiation

Cited by 166 publications

References 46 publications

β-Catenin induces immortalization of melanocytes by suppressing p16INK4a expression and cooperates with N-Ras in melanoma development

β-Catenin induces immortalization of melanocytes by suppressing p16INK4a expression and cooperates with N-Ras in melanoma development

BLOC-1 Is Required for Cargo-specific Sorting from Vacuolar Early Endosomes toward Lysosome-related Organelles

Isolation, Culture, and Transfection of Melanocytes

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β-Catenin induces immortalization of melanocytes by suppressing p16^INK4a expression and cooperates with N-Ras in melanoma development

β-Catenin induces immortalization of melanocytes by suppressing p16^INK4a expression and cooperates with N-Ras in melanoma development