Abstract
Altered expression of human Scribble is associated with invasive epithelial cancers, however, its role in tumour development remains unclear. Mutations in Drosophila Scribble result in loss of polarity, overproliferation and 3D-tumourous overgrowth of epithelial cells. Using complementation studies in Drosophila we recently demonstrated that expression of human Scribble can also regulate polarity and restrict tissue overgrowth. Here, we have undertaken a detailed study of human Scribble function in the polarized mammary cell line, MCF10A. We show that although Scribble does not seem to be required for apical-basal polarity or proliferation control in MCF10A cells, Scribble is essential for the control of polarity associated with directed epithelial cell migration. Scribble-depleted MCF10A cells show defective in vitro wound closure and chemotactic movement. The cells at the wound edge fail to polarize, show reduced lamellipodia formation and impaired recruitment of Cdc42 and Rac1 to the leading edge. Furthermore, we show that this function is relevant in vivo as Scribble mutant mice show defective epidermal wound healing. This data identifies an essential role for mammalian Scribble in the regulation of the polarity specifically involved in directed epithelial migration.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 50 print issues and online access
$259.00 per year
only $5.18 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
Abbreviations
- MTOC:
-
microtubule organizing centre
References
Audebert S, Navarro C, Nourry C, Chasserot-Golaz S, Lecine P, Bellaiche Y et al. (2004). Mammalian Scribble forms a tight complex with the betaPIX exchange factor. Curr Biol 14: 987–995.
Berkowitz EA, Seroogy KB, Schroeder JA, Russell WE, Evans EP, Riedel RF et al. (1996). Characterization of the mouse transforming growth factor alpha gene: its expression during eyelid development and in waved 1 tissues. Cell Growth Differ 7: 1271–1282.
Bilder D . (2004). Epithelial polarity and proliferation control: links from the Drosophila neoplastic tumor suppressors. Genes Dev 18: 1909–1925.
Bilder D, Perrimon N . (2000). Localization of apical epithelial determinants by the basolateral PDZ protein Scribble. Nature 403: 676–680.
Bilder D, Schober M, Perrimon N . (2003). Integrated activity of PDZ protein complexes regulates epithelial polarity. Nat Cell Biol 5: 53–58.
Boussioutas A, Li H, Liu J, Waring P, Lade S, Holloway AJ et al. (2003). Distinctive patterns of gene expression in premalignant gastric mucosa and gastric cancer. Cancer Res 63: 2569–2577.
Brumby AM, Richardson HE . (2003). scribble mutants cooperate with oncogenic Ras or Notch to cause neoplastic overgrowth in Drosophila. EMBO J 22: 5769–5779.
Brummelkamp TR, Bernards R, Agami R . (2002). Stable suppression of tumorigenicity by virus-mediated RNA interference. Cancer Cell 2: 243–247.
Cau J, Hall A . (2005). Cdc42 controls the polarity of the actin and microtubule cytoskeletons through two distinct signal transduction pathways. J Cell Sci 118: 2579–2587.
Cavatorta AL, Fumero G, Chouhy D, Aguirre R, Nocito AL, Giri AA et al. (2004). Differential expression of the human homologue of Drosophila discs large oncosuppressor in histologic samples from human papillomavirus-associated lesions as a marker for progression to malignancy. Int J Cancer 111: 373–380.
Chen X, Macara IG . (2005). Par-3 controls tight junction assembly through the Rac exchange factor Tiam1. Nat Cell Biol 7: 262–269.
Debnath J, Muthuswamy SK, Brugge JS . (2003). Morphogenesis and oncogenesis of MCF-10A mammary epithelial acini grown in three-dimensional basement membrane cultures. Methods 30: 256–268.
Dow LE, Brumby AM, Muratore R, Coombe ML, Sedelies KA, Trapani JA et al. (2003). hScrib is a functional homologue of the Drosophila tumour suppressor Scribble. Oncogene 22: 9225–9230.
Etienne-Manneville S, Hall A . (2001). Integrin-mediated activation of Cdc42 controls cell polarity in migrating astrocytes through PKCzeta. Cell 106: 489–498.
Etienne-Manneville S, Hall A . (2003). Cdc42 regulates GSK-3beta and adenomatous polyposis coli to control cell polarity. Nature 421: 753–756.
Etienne-Manneville S, Manneville JB, Nicholls S, Ferenczi MA, Hall A . (2005). Cdc42 and Par6-PKCzeta regulate the spatially localized association of Dlg1 and APC to control cell polarization. J Cell Biol 170: 895–901.
Gardiol D, Zacchi A, Petrera F, Stanta G, Banks L . (2006). Human discs large and scrib are localized at the same regions in colon mucosa and changes in their expression patterns are correlated with loss of tissue architecture during malignant progression. Int J Cancer 119: 1285–1290.
Gomes ER, Jani S, Gundersen GG . (2005). Nuclear movement regulated by Cdc42, MRCK, myosin, and actin flow establishes MTOC polarization in migrating cells. Cell 121: 451–463.
Hall A . (2005). Rho GTPases and the control of cell behaviour. Biochem Soc Trans 33: 891–895.
Humbert P, Russell S, Richardson H . (2003). Dlg, Scribble and Lgl in cell polarity, cell proliferation and cancer. Bioessays 25: 542–553.
Humbert PO, Dow LE, Russell SM . (2006). The Scribble and Par complexes in polarity and migration: Friends or Foes? Trends Cell Biol, in press.
Legouis R, Gansmuller A, Sookhareea S, Bosher JM, Baillie DL, Labouesse M . (2000). LET-413 is a basolateral protein required for the assembly of adherens junctions in Caenorhabditis elegans. Nat Cell Biol 2: 415–422.
Legouis R, Jaulin-Bastard F, Schott S, Navarro C, Borg JP, Labouesse M . (2003). Basolateral targeting by leucine-rich repeat domains in epithelial cells. EMBO Rep 4: 1096–1102.
Lin HT, Steller MA, Aish L, Hanada T, Chishti AH . (2004). Differential expression of human Dlg in cervical intraepithelial neoplasias. Gynecol Oncol 93: 422–428.
Ludford-Menting MJ, Oliaro J, Sacirbegovic F, Cheah ET, Pedersen N, Thomas SJ et al. (2005). A network of PDZ-containing proteins regulates T cell polarity and morphology during migration and immunological synapse formation. Immunity 22: 737–748.
Massimi P, Gammoh N, Thomas M, Banks L . (2004). HPV E6 specifically targets different cellular pools of its PDZ domain-containing tumour suppressor substrates for proteasome-mediated degradation. Oncogene 23: 8033–8039.
McCluskey J, Hopkinson-Woolley J, Luke B, Martin P . (1993). A study of wound healing in the E11.5 mouse embryo by light and electron microscopy. Tissue Cell 25: 173–181.
Mine N, Iwamoto R, Mekada E . (2005). HB-EGF promotes epithelial cell migration in eyelid development. Development 132: 4317–4326.
Murdoch JN, Henderson DJ, Doudney K, Gaston-Massuet C, Phillips HM, Paternotte C et al. (2003). Disruption of scribble (Scrb1) causes severe neural tube defects in the circletail mouse. Hum Mol Genet 12: 87–98.
Nakagawa S, Huibregtse JM . (2000). Human scribble (Vartul) is targeted for ubiquitin-mediated degradation by the high-risk papillomavirus E6 proteins and the E6AP ubiquitin-protein ligase. Mol Cell Biol 20: 8244–8253.
Nakagawa S, Yano T, Nakagawa K, Takizawa S, Suzuki Y, Yasugi T et al. (2004). Analysis of the expression and localisation of a LAP protein, human scribble, in the normal and neoplastic epithelium of uterine cervix. Br J Cancer 90: 194–199.
Navarro C, Nola S, Audebert S, Santoni MJ, Arsanto JP, Ginestier C et al. (2005). Junctional recruitment of mammalian Scribble relies on E-cadherin engagement. Oncogene 24: 4330–4339.
Nishiya N, Kiosses WB, Han J, Ginsberg MH . (2005). An alpha4 integrin-paxillin-Arf-GAP complex restricts Rac activation to the leading edge of migrating cells. Nat Cell Biol 7: 343–352.
Nobes CD, Hall A . (1999). Rho GTPases control polarity, protrusion, and adhesion during cell movement. J Cell Biol 144: 1235–1244.
Ozdamar B, Bose R, Barrios-Rodiles M, Wang HR, Zhang Y, Wrana JL . (2005). Regulation of the polarity protein Par6 by TGFbeta receptors controls epithelial cell plasticity. Science 307: 1603–1609.
Pagliarini RA, Xu T . (2003). A genetic screen in Drosophila for metastatic behavior. Science 302: 1227–1231.
Petersen OW, Ronnov-Jessen L, Howlett AR, Bissell MJ . (1992). Interaction with basement membrane serves to rapidly distinguish growth and differentiation pattern of normal and malignant human breast epithelial cells. Proc Natl Acad Sci USA 89: 9064–9068.
Pinheiro EM, Montell DJ . (2004). Requirement for Par-6 and Bazooka in Drosophila border cell migration. Development 131: 5243–5251.
Qin Y, Capaldo C, Gumbiner BM, Macara IG . (2005). The mammalian Scribble polarity protein regulates epithelial cell adhesion and migration through E-cadherin. J Cell Biol 171: 1061–1071.
Redvers RP, Kaur P . (2005). Serial cultivation of primary adult murine keratinocytes. Methods Mol Biol 289: 15–22.
Shin I, Kim S, Song H, Kim HR, Moon A . (2005). H-Ras-specific activation of Rac-MKK3/6-p38 pathway: its critical role in invasion and migration of breast epithelial cells. J Biol Chem 280: 14675–14683.
Thomas M, Massimi P, Navarro C, Borg JP, Banks L . (2005). The hScrib/Dlg apico-basal control complex is differentially targeted by HPV-16 and HPV-18 E6 proteins. Oncogene 24: 6222–6230.
Ting SB, Caddy J, Hislop N, Wilanowski T, Auden A, Zhao LL et al. (2005). A homolog of Drosophila grainy head is essential for epidermal integrity in mice. Science 308: 411–413.
Wada H, Iwasaki M, Sato T, Masai I, Nishiwaki Y, Tanaka H et al. (2005). Dual roles of zygotic and maternal Scribble1 in neural migration and convergent extension movements in zebrafish embryos. Development 132: 2273–2285.
Watanabe T, Wang S, Noritake J, Sato K, Fukata M, Takefuji M et al. (2004). Interaction with IQGAP1 links APC to Rac1, Cdc42, and actin filaments during cell polarization and migration. Dev Cell 7: 871–883.
Watson RA, Rollason TP, Reynolds GM, Murray PG, Banks L, Roberts S . (2002). Changes in expression of the human homologue of the Drosophila discs large tumour suppressor protein in high-grade premalignant cervical neoplasias. Carcinogenesis 23: 1791–1796.
Zarbalis K, May SR, Shen Y, Ekker M, Rubenstein JL, Peterson AS . (2004). A focused and efficient genetic screening strategy in the mouse: identification of mutations that disrupt cortical development. PLoS Biol 2: E219.
Zenz R, Scheuch H, Martin P, Frank C, Eferl R, Kenner L et al. (2003). c-Jun regulates eyelid closure and skin tumor development through EGFR signaling. Dev Cell 4: 879–889.
Acknowledgements
We would like to thank members of the Bissell lab, and J Debnath and S Muthuswamy for advice with the culture of MCF10A cells. We thank S Ellis and N Waterhouse for assistance with confocal and live-cell microscopy, R Redvers for advice on culture of primary keratinocytes and J Hayes for maintenance of the scribblerumz mouse colony. We thank H Richardson and A Brumby for critical reading of the manuscript. This work was supported by project grants from the National Institutes of Health (USA) (PO1 HL53749⁁03) and Australian National Health and Medical Research Council (282400) (SMJ), the Australian Research Council (SMR), the Association for International Cancer Research UK and The Cancer Council Victoria (POH, SMR). LED was supported by a Cancer Council Victoria Postgraduate Cancer Research Scholarship, JSK was supported by The LAVFW Postdoctoral Research Fellowship and POH was supported by a Career Development Award from the Australian National Health and Medical Research Council.
Author information
Authors and Affiliations
Corresponding author
Additional information
Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc).
Rights and permissions
About this article
Cite this article
Dow, L., Kauffman, J., Caddy, J. et al. The tumour-suppressor Scribble dictates cell polarity during directed epithelial migration: regulation of Rho GTPase recruitment to the leading edge. Oncogene 26, 2272–2282 (2007). https://doi.org/10.1038/sj.onc.1210016
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/sj.onc.1210016
Keywords
This article is cited by
-
Micronutrient supplementation affects DNA methylation in male gonads with potential intergenerational epigenetic inheritance involving the embryonic development through glutamate receptor-associated genes
BMC Genomics (2022)
-
TSPAN6 is a suppressor of Ras-driven cancer
Oncogene (2022)
-
The Scribble family in cancer: twentieth anniversary
Oncogene (2020)
-
Diverging impact of cell fate determinants Scrib and Llgl1 on adhesion and migration of hematopoietic stem cells
Journal of Cancer Research and Clinical Oncology (2018)
-
TAp63 suppresses mammary tumorigenesis through regulation of the Hippo pathway
Oncogene (2017)