Abstract
The specific ablation of Rb1 gene in epidermis (RbF/F;K14cre) promotes proliferation and altered differentiation but does not produce spontaneous tumour development. These phenotypic changes are associated with increased expression of E2F members and E2F-dependent transcriptional activity. Here, we have focused on the possible dependence on E2F1 gene function. We have generated mice that lack Rb1 in epidermis in an inducible manner (RbF/F;K14creERTM). These mice are indistinguishable from those lacking pRb in this tissue in a constitutive manner (RbF/F;K14cre). In an E2F1-null background (RbF/F;K14creERTM; and E2F1−/− mice), the phenotype due to acute Rb1 loss is not ameliorated by E2F1 loss, but rather exacerbated, indicating that pRb functions in epidermis do not rely solely on E2F1. On the other hand, RbF/F;K14creERTM;E2F1−/− mice develop spontaneous epidermal tumours of hair follicle origin with high incidence. These tumours, which retain a functional p19arf/p53 axis, also show aberrant activation of β-catenin/Wnt pathway. Gene expression studies revealed that these tumours display relevant similarities with specific human tumours. These data demonstrate that the Rb/E2F1 axis exerts essential functions not only in maintaining epidermal homoeostasis, but also in suppressing tumour development in epidermis, and that the disruption of this pathway may induce tumour progression through specific alteration of developmental programs.
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References
Nevins JR . The Rb/E2F pathway and cancer. Hum Mol Genet 2001; 10: 699–703.
Ruiz S, Santos M, Segrelles C, Leis H, Jorcano JL, Berns A et al. Unique and overlapping functions of pRb and p107 in the control of proliferation and differentiation in epidermis. Development 2004; 131: 2737–2748.
Ruiz S, Santos M, Lara MF, Segrelles C, Ballestin C, Paramio JM . Unexpected roles for pRb in mouse skin carcinogenesis. Cancer Res 2005; 65: 9678–9686.
Martinez-Cruz AB, Santos M, Lara MF, Segrelles C, Ruiz S, Moral M et al. Spontaneous squamous cell carcinoma induced by the somatic inactivation of retinoblastoma and Trp53 tumor suppressors. Cancer Res 2008; 68: 683–692.
Lara MF, Santos M, Ruiz S, Segrelles C, Moral M, Martinez-Cruz AB et al. p107 acts as a tumor suppressor in pRb-deficient epidermis. Mol Carcinog 2008; 47: 105–113.
Santos M, Ruiz S, Lara MF, Segrelles C, Moral M, Martinez-Cruz AB et al. Susceptibility of pRb-deficient epidermis to chemical skin carcinogenesis is dependent on the p107 allele dosage. Mol Carcinog 2008; 47: 815–821.
Sage J, Miller AL, Perez-Mancera PA, Wysocki JM, Jacks T . Acute mutation of retinoblastoma gene function is sufficient for cell cycle re-entry. Nature 2003; 424: 223–228.
Johnson DG, Cress WD, Jakoi L, Nevins JR . Oncogenic capacity of the E2F1 gene. Proc Nat Acad Sci USA 1994; 91: 12823–12827.
Johnson DG, Schwarz JK, Cress WD, Nevins JR . Expression of transcription factor E2F1 induces quiescent cells to enter S phase. Nature 1993; 365: 349–352.
Field SJ, Tsai FY, Kuo F, Zubiaga AM, Kaelin WG, Livingston DM et al. E2F-1 functions in mice to promote apoptosis and suppress proliferation. Cell 1996; 85: 549–561.
Yamasaki L, Jacks T, Bronson R, Goillot E, Harlow E, Dyson NJ . Tumor induction and tissue atrophy in mice lacking E2F-1. Cell 1996; 85: 537–548.
Yamasaki L, Bronson R, Williams BO, Dyson NJ, Harlow E, Jacks T . Loss of E2F-1 reduces tumorigenesis and extends the lifespan of Rb1(+/-)mice. Nature genetics 1998; 18: 360–364.
Tsai KY, Hu Y, Macleod KF, Crowley D, Yamasaki L, Jacks T . Mutation of E2f-1 suppresses apoptosis and inappropriate S phase entry and extends survival of Rb-deficient mouse embryos. Molecular cell 1998; 2: 293–304.
Paramio JM, Segrelles C, Casanova ML, Jorcano JL . Opposite functions for E2F1 and E2F4 in human epidermal keratinocyte differentiation. J Biol Chem 2000; 275: 41219–41226.
D'Souza SJ, Vespa A, Murkherjee S, Maher A, Pajak A, Dagnino L . E2F-1 is essential for normal epidermal wound repair. J Biol Chem 2002; 277 (): 10626–10632.
Lorz C, Garcia-Escudero R, Segrelles C, Garin MI, Ariza JM, Santos M et al. A functional role of RB-dependent pathway in the control of quiescence in adult epidermal stem cells revealed by genomic profiling. Stem cell reviews 2010; 6: 162–177.
Lopez RG, Garcia-Silva S, Moore SJ, Bereshchenko O, Martinez-Cruz AB, Ermakova O et al. C/EBPalpha and beta couple interfollicular keratinocyte proliferation arrest to commitment and terminal differentiation. Nat Cell Biol 2009; 11: 1181–1190.
Pierce AM, Fisher SM, Conti CJ, Johnson DG . Deregulated expression of E2F1 induces hyperplasia and cooperates with ras in skin tumor development. Oncogene 1998; 16: 1267–1276.
Pierce AM, Gimenez-Conti IB, Schneider-Broussard R, Martinez LA, Conti CJ, Johnson DG . Increased E2F1 activity induces skin tumors in mice heterozygous and nullizygous for p53. Proc Nat Acad Sci USA 1998; 95: 8858–8863.
Pierce AM, Schneider-Broussard R, Gimenez-Conti IB, Russell JL, Conti CJ, Johnson DG . E2F1 has both oncogenic and tumor-suppressive properties in a transgenic model. Mol Cell Biol 1999; 19: 6408–6414.
Wikonkal NM, Remenyik E, Knezevic D, Zhang W, Liu M, Zhao H et al. Inactivating E2f1 reverts apoptosis resistance and cancer sensitivity in Trp53-deficient mice. Nat Cell Biol 2003; 5: 655–660.
Russell JL, Weaks RL, Berton TR, Johnson DG . E2F1 suppresses skin carcinogenesis via the ARF-p53 pathway. Oncogene 2006; 25: 867–876.
Lara MF, Garcia-Escudero R, Ruiz S, Santos M, Moral M, Martinez-Cruz AB et al. Gene profiling approaches help to define the specific functions of retinoblastoma family in epidermis. Mol Carcinog 2008; 47: 209–221.
Lachmann A, Xu H, Krishnan J, Berger SI, Mazloom AR, Ma'ayan A . ChEA: transcription factor regulation inferred from integrating genome-wide ChIP-X experiments. Bioinformatics 2010; 26: 2438–2444.
O'Guin WM, Sun TT, Manabe M . Interaction of trichohyalin with intermediate filaments: three immunologically defined stages of trichohyalin maturation. J Invest Dermatol 1992; 98: 24–32.
Ruiz S, Santos M, Paramio JM . Is the loss of pRb essential for the mouse skin carcinogenesis? Cell Cycle 2006; 5: 625–629.
Morris EJ, Ji JY, Yang F, Di Stefano L, Herr A, Moon NS et al. E2F1 represses beta-catenin transcription and is antagonized by both pRB and CDK8. Nature 2008; 455: 552–556.
Gat U, DasGupta R, Degenstein L, Fuchs E . De Novo hair follicle morphogenesis and hair tumors in mice expressing a truncated beta-catenin in skin. Cell 1998; 95: 605–614.
Lo Celso C, Prowse DM, Watt FM . Transient activation of beta-catenin signalling in adult mouse epidermis is sufficient to induce new hair follicles but continuous activation is required to maintain hair follicle tumours. Development 2004; 131: 1787–1799.
Doglioni C, Piccinin S, Demontis S, Cangi MG, Pecciarini L, Chiarelli C et al. Alterations of beta-catenin pathway in non-melanoma skin tumors: loss of alpha-ABC nuclear reactivity correlates with the presence of beta-catenin gene mutation. Am J pathol 2003; 163: 2277–2287.
Lowry WE, Blanpain C, Nowak JA, Guasch G, Lewis L, Fuchs E . Defining the impact of beta-catenin/Tcf transactivation on epithelial stem cells. Genes Dev 2005; 19: 1596–1611.
Huelsken J, Vogel R, Erdmann B, Cotsarelis G, Birchmeier W . beta-Catenin controls hair follicle morphogenesis and stem cell differentiation in the skin. Cell 2001; 105: 533–545.
Li M, He Y, Dubois W, Wu X, Shi J, Huang J . Distinct regulatory mechanisms and functions for p53-activated and p53-repressed DNA damage response genes in embryonic stem Cells. Molecular cell 2012; 46: 30–42.
Rhodes DR, Kalyana-Sundaram S, Mahavisno V, Varambally R, Yu J, Briggs BB et al. Oncomine 3.0: genes, pathways, and networks in a collection of 18,000 cancer gene expression profiles. Neoplasia 2007; 9: 166–180.
Rhodes DR, Yu J, Shanker K, Deshpande N, Varambally R, Ghosh D et al. ONCOMINE: a cancer microarray database and integrated data-mining platform. Neoplasia 2004; 6: 1–6.
Chong JL, Wenzel PL, Saenz-Robles MT, Nair V, Ferrey A, Hagan JP et al. E2f1-3 switch from activators in progenitor cells to repressors in differentiating cells. Nature 2009; 462: 930–934.
Chong JL, Tsai SY, Sharma N, Opavsky R, Price R, Wu L et al. E2f3a and E2f3b contribute to the control of cell proliferation and mouse development. Mol Cell Biol 2009; 29: 414–424.
Saavedra HI, Wu L, de Bruin A, Timmers C, Rosol TJ, Weinstein M et al. Specificity of E2F1, E2F2, and E2F3 in mediating phenotypes induced by loss of Rb. Cell Growth Differ 2002; 13: 215–225.
Rabbani F, Cordon-Cardo C . Mutation of cell cycle regulators and their impact on superficial bladder cancer. Urol clin North Am 2000; 27: 83–102 ix.
Rabbani F, Richon VM, Orlow I, Lu ML, Drobnjak M, Dudas M et al. Prognostic significance of transcription factor E2F-1 in bladder cancer: genotypic and phenotypic characterization. J Natl Cancer Ins 1999; 91: 874–881.
Lee JS, Leem SH, Lee SY, Kim SC, Park ES, Kim SB et al. Expression signature of E2F1 and its associated genes predict superficial to invasive progression of bladder tumors. J Clin Oncol 2010; 28: 2660–2667.
Rounbehler RJ, Rogers PM, Conti CJ, Johnson DG . Inactivation of E2f1 enhances tumorigenesis in a Myc transgenic model. Cancer Res 2002; 62: 3276–3281.
Paulson QX, McArthur MJ, Johnson DG . E2F3a stimulates proliferation, p53-independent apoptosis and carcinogenesis in a Transgenic Mouse Model. Cell Cycle 2006; 5: 184–190.
Ziebold U, Reza T, Caron A, Lees JA . E2F3 contributes both to the inappropriate proliferation and to the apoptosis arising in Rb mutant embryos. Genes Dev 2001; 15: 386–391.
Ruiz S, Segrelles C, Santos M, Lara MF, Paramio JM . Functional link between retinoblastoma family of proteins and the Wnt signaling pathway in mouse epidermis. Dev Dyn 2004; 230: 410–418.
Takebe N, Harris PJ, Warren RQ, Ivy SP . Targeting cancer stem cells by inhibiting Wnt, Notch, and Hedgehog pathways. Nat Rev Clin Oncol 2011; 8: 97–106.
Lobo NA, Shimono Y, Qian D, Clarke MF . The biology of cancer stem cells. Annu Rev Cell Dev Biol 2007; 23: 675–699.
Clevers H . Wnt/beta-catenin signaling in development and disease. Cell 2006; 127: 469–480.
Chan EF, Gat U, McNiff JM, Fuchs E . A common human skin tumour is caused by activating mutations in beta-catenin. Nature genetics 1999; 21: 410–413.
Gandarillas A, Watt FM . c-Myc promotes differentiation of human epidermal stem cells. Genes Dev 1997; 11: 2869–2882.
Haegebarth A, Clevers H . Wnt signaling, lgr5, and stem cells in the intestine and skin. Am J pathol 2009; 174: 715–721.
Alonso L, Fuchs E . Stem cells in the skin: waste not, Wnt not. Genes Dev 2003; 17: 1189–1200.
Lorz C, Segrelles C, Paramio JM . On the origin of epidermal cancers. Curr Mol med 2009; 9: 353–364.
Ahmad I, Sansom OJ, Leung HY . Exploring molecular genetics of bladder cancer: lessons learned from mouse models. Dis models mech 2012; 5: 323–332.
Ahmad I, Patel R, Liu Y, Singh LB, Taketo MM, Wu XR et al. Ras mutation cooperates with beta-catenin activation to drive bladder tumourigenesis. Cell death disease 2011; 2: e124.
Ahmad I, Morton JP, Singh LB, Radulescu SM, Ridgway RA, Patel S et al. beta-Catenin activation synergizes with PTEN loss to cause bladder cancer formation. Oncogene 2011; 30: 178–189.
Shin K, Lee J, Guo N, Kim J, Lim A, Qu L et al. Hedgehog/Wnt feedback supports regenerative proliferation of epithelial stem cells in bladder. Nature 2011; 472: 110–114.
Brandt WD, Matsui W, Rosenberg JE, He X, Ling S, Schaeffer EM et al. Urothelial carcinoma: stem cells on the edge. Cancer metastasis rev 2009; 28: 291–304.
Segrelles C, Lu J, Hammann B, Santos M, Moral M, Cascallana JL et al. Deregulated activity of akt in epithelial basal cells induces spontaneous tumors and heightened sensitivity to skin carcinogenesis. Cancer Res 2007; 67: 10879–10888.
Paramio JM, Casanova ML, Segrelles C, Mittnacht S, Lane EB, Jorcano JL . Modulation of cell proliferation by cytokeratins K10 and K16. Mol Cell Biol 1999; 19: 3086–3094.
Segrelles C, Moral M, Lorz C, Santos M, Lu J, Cascallana JL et al. Constitutively active akt induces ectodermal defects and impaired bone morphogenetic protein signaling. Mol biol cell 2008; 19: 137–149.
Lorz C, Segrelles C, Garin M, Paramio JM . Isolation of adult mouse stem keratinocytes using magnetic cell sorting (MACS). Methods in mol biol 2010; 585: 1–11.
Saeed AI, Sharov V, White J, Li J, Liang W, Bhagabati N et al. TM4: a free, open-source system for microarray data management and analysis. Biotechniques 2003; 34: 374–378.
Chen R, Li L, Butte AJ . AILUN: reannotating gene expression data automatically. Nature methods 2007; 4: 879.
Dennis G, Sherman BT, Hosack DA, Yang J, Gao W, Lane HC et al. DAVID: Database for Annotation, Visualization, and Integrated Discovery. Genome Biol 2003; 4: P3.
Hosack DA, Dennis G, Sherman BT, Lane HC, Lempicki RA . Identifying biological themes within lists of genes with EASE. Genome Biol 2003; 4: R70.
Subramanian A, Tamayo P, Mootha VK, Mukherjee S, Ebert BL, Gillette MA et al. Gene set enrichment analysis: a knowledge-based approach for interpreting genome-wide expression profiles. Proc Natl Acad Sci USA 2005; 102: 15545–15550.
Acknowledgements
Ministerio de Ciencia e Innovación (MICINN) grants SAF2006–0121, SAF2011–26122-C02–01 and JCI-2010–06167, Comunidad Autónoma de Madrid Oncocycle Program Grant S2006/BIO-0232 and: CAM P2010/BMD-2470, Ministerio de Sanidad y Consumo grant ISCIII-RETIC RD06/0020/0029 and from Fundación Sandra Ibarra to JMP. The excellent technical support by Pilar Hernández in histology and the personnel of the CIEMAT Animal Facility are specially acknowledged.
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Costa, C., Santos, M., Martínez-Fernández, M. et al. E2F1 loss induces spontaneous tumour development in Rb-deficient epidermis. Oncogene 32, 2937–2951 (2013). https://doi.org/10.1038/onc.2012.316
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DOI: https://doi.org/10.1038/onc.2012.316
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