Nhận diện các oncogen mới hợp tác với Ras trong Drosophila melanogaster: Đường dẫn RhoGEF/Rho-Family/JNK là động lực chính của quá trình ung thư hóa
Tóm tắt
Chúng tôi đã chỉ ra trước đây rằng các đột biến trong các yếu tố điều chỉnh tính phân cực tế bào apico-basal hợp tác với Ras oncogenic (RasACT) để thúc đẩy quá trình ung thư ở Drosophila melanogaster và tế bào động vật có vú. Để xác định các gen mới hợp tác với RasACT trong quá trình ung thư hóa, chúng tôi đã thực hiện sàng lọc toàn bộ hệ gen để tìm các gen mà khi được biểu hiện quá mức trong mắt Drosophila đang phát triển sẽ tăng cường sự phì đại do RasACT thúc đẩy. Các gen hợp tác với RasACT được xác định là Rac1, Rho1, RhoGEF2, pbl, rib, và east, những gen này mã hóa cho các yếu tố điều chỉnh hình thái tế bào. Trong một bối cảnh đồng phân (clonal setting), điều này cho thấy các gen mang lại lợi thế cạnh tranh so với tế bào hoang dã, chỉ Rac1, một alen hoạt hóa của Rho1 (Rho1ACT), RhoGEF2, và pbl đã hợp tác với RasACT, dẫn đến giảm sự phân hóa và tạo thành các khối u xâm lấn lớn. Sự biểu hiện của RhoGEF2 hoặc Rac1 cùng với RasACT đã tăng cường hoạt động của Jun kinase (JNK), và việc tăng cường JNK là cần thiết cho sự hợp tác. Tuy nhiên, trong hệ thống toàn bộ mô, việc tăng cường JNK một mình không đủ để hợp tác với RasACT, trong khi trong bối cảnh đồng phân, việc tăng cường JNK là đủ cho quá trình ung thư hóa do RasACT điều khiển. Sự tăng cường JNK cũng đủ để tạo ra sự phát triển xâm lấn của các tế bào biểu mô vú MCF10A biểu hiện RasV12. Nhất quán với điều này, các khối u vú ở người HER2+ (nơi mà yếu tố tăng trưởng biểu bì người 2 được biểu hiện quá mức và tín hiệu Ras được tăng cường) cho thấy một mối tương quan đáng kể với chữ ký biểu thị hoạt hóa đường dẫn JNK. Hơn nữa, phân tích di truyền của chúng tôi ở Drosophila đã chỉ ra rằng Rho1 và Rac đóng vai trò quan trọng trong việc hợp tác của sự biểu hiện quá mức RhoGEF2 hoặc Pbl và các đột biến trong các yếu tố điều chỉnh tính phân cực, Dlg và aPKC, với RasACT trong bối cảnh toàn bộ mô. Tổng thể, phân tích của chúng tôi cho thấy tầm quan trọng của đường dẫn RhoGEF/Rho-family/JNK trong quá trình ung thư hóa hợp tác với RasACT.
Từ khóa
Tài liệu tham khảo
Adachi-Yamada, 2004, Mechanisms for removal of developmentally abnormal cells: cell competition and morphogenetic apoptosis, J. Biochem., 136, 13, 10.1093/jb/mvh099
Aigaki, 2001, The gene search system: its application to functional genomics in Drosophila melanogaster, J. Neurogenet., 15, 169, 10.3109/01677060109167374
Arias-Romero, 2010, A Rac–Pak signaling pathway is essential for ErbB2-mediated transformation of human breast epithelial cancer cells, Oncogene, 29, 5839, 10.1038/onc.2010.318
Baonza, 2005, Control of cell proliferation in the Drosophila eye by Notch signaling, Dev. Cell, 8, 529, 10.1016/j.devcel.2005.01.019
Barrett, 1997, The Rho GTPase and a putative RhoGEF mediate a signaling pathway for the cell shape changes in Drosophila gastrulation, Cell, 91, 905, 10.1016/S0092-8674(00)80482-1
Beaucher, 2006, Metastatic ability of Drosophila tumors depends on MMP activity, Dev. Biol., 303, 625, 10.1016/j.ydbio.2006.12.001
Bergantinos, 2010, Cell death-induced regeneration in wing imaginal discs requires JNK signalling, Development, 137, 1169, 10.1242/dev.045559
Bergmann, 1998, The Drosophila gene hid is a direct molecular target of Ras-dependent survival signaling, Cell, 95, 331, 10.1016/S0092-8674(00)81765-1
Betschinger, 2003, The Par complex directs asymmetric cell division by phosphorylating the cytoskeletal protein Lgl, Nature, 422, 326, 10.1038/nature01486
Bilder, 2000, Localization of apical epithelial determinants by the basolateral PDZ protein Scribble, Nature, 403, 676, 10.1038/35001108
Bilder, 2000, Cooperative regulation of cell polarity and growth by Drosophila tumor suppressors, Science, 289, 113, 10.1126/science.289.5476.113
Billuart, 2001, Regulating axon branch stability: the role of p190 RhoGAP in repressing a retraction signaling pathway, Cell, 107, 195, 10.1016/S0092-8674(01)00522-0
Blake, 1998, The products of ribbon and raw are necessary for proper cell shape and cellular localization of nonmuscle myosin in Drosophila, Dev. Biol., 203, 177, 10.1006/dbio.1998.9036
Bradley, 2001, ribbon encodes a novel BTB/POZ protein required for directed cell migration in Drosophila melanogaster, Development, 128, 3001, 10.1242/dev.128.15.3001
Brumby, 2003, scribble mutants cooperate with oncogenic Ras or Notch to cause neoplastic overgrowth in Drosophila, EMBO J., 22, 5769, 10.1093/emboj/cdg548
Brumby, 2005, Using Drosophila melanogaster to map human cancer pathways, Nat. Rev. Cancer, 5, 626, 10.1038/nrc1671
Ciapponi, 2001, Drosophila Fos mediates ERK and JNK signals via distinct phosphorylation sites, Genes Dev., 15, 1540, 10.1101/gad.886301
Coleman, 2004, RAS and RHO GTPases in G1-phase cell-cycle regulation, Nat. Rev. Mol. Cell Biol., 5, 355, 10.1038/nrm1365
Cordero, 2010, Oncogenic Ras diverts a host TNF tumor suppressor activity into tumor promoter, Dev. Cell, 18, 999, 10.1016/j.devcel.2010.05.014
Debnath, 2003, Morphogenesis and oncogenesis of MCF-10A mammary epithelial acini grown in three-dimensional basement membrane cultures, Methods, 30, 256, 10.1016/S1046-2023(03)00032-X
DeNicola, 2009, RAS in cellular transformation and senescence, Eur. J. Cancer, 45, 211, 10.1016/S0959-8049(09)70036-X
Dietzl, 2007, A genome-wide transgenic RNAi library for conditional gene inactivation in Drosophila, Nature, 448, 151, 10.1038/nature05954
Dow, 2008, Loss of human Scribble cooperates with H-Ras to promote cell invasion through deregulation of MAPK signalling, Oncogene, 27, 5988, 10.1038/onc.2008.219
Fan, 2008, Apoptosis-induced compensatory proliferation: the Cell is dead. Long live the Cell!, Trends Cell Biol., 18, 467, 10.1016/j.tcb.2008.08.001
Ferres-Marco, 2006, Epigenetic silencers and Notch collaborate to promote malignant tumours by Rb silencing, Nature, 439, 430, 10.1038/nature04376
Froldi, 2010, The lethal giant larvae tumour suppressor mutation requires dMyc oncoprotein to promote clonal malignancy, BMC Biol., 8, 33, 10.1186/1741-7007-8-33
Fukuhara, 1999, A novel PDZ domain containing guanine nucleotide exchange factor links heterotrimeric G proteins to Rho, J. Biol. Chem., 274, 5868, 10.1074/jbc.274.9.5868
Fukuhara, 2000, Leukemia-associated Rho guanine nucleotide exchange factor (LARG) links heterotrimeric G proteins of the G(12) family to Rho, FEBS Lett., 485, 183, 10.1016/S0014-5793(00)02224-9
Fukuhara, 2001, RGS-containing RhoGEFs: The missing link between transforming G proteins and Rho?, Oncogene, 20, 1661, 10.1038/sj.onc.1204182
Garlena, 2010, Regulation of mixed-lineage kinase activation in JNK-dependent morphogenesis, J. Cell Sci., 123, 3177, 10.1242/jcs.063313
Gateff, 1978, Malignant neoplasms of genetic origin in Drosophila melanogaster, Science, 200, 1448, 10.1126/science.96525
Gateff, 1969, Neoplasms in mutant and cultured wild-tupe tissues of Drosophila, Natl. Cancer Inst. Monogr., 31, 365
Gomez del Pulgar, 2005, Rho GTPase expression in tumourigenesis: evidence for a significant link, BioEssays, 27, 602, 10.1002/bies.20238
Greco, 2001, Argosomes: a potential vehicle for the spread of morphogens through epithelia, Cell, 106, 633, 10.1016/S0092-8674(01)00484-6
Grusche, 2011, The Salvador/Warts/Hippo pathway controls regenerative tissue growth in Drosophila melanogaster, Dev. Biol., 350, 255, 10.1016/j.ydbio.2010.11.020
Grzeschik, 2007, Abnormalities in cell proliferation and apico-basal cell polarity are separable in Drosophila lgl mutant clones in the developing eye, Dev. Biol., 311, 106, 10.1016/j.ydbio.2007.08.025
Grzeschik, 2010, Lgl, aPKC, and Crumbs regulate the Salvador/Warts/Hippo pathway through two distinct mechanisms, Curr. Biol., 20, 573, 10.1016/j.cub.2010.01.055
Hacker, 1998, DRhoGEF2 encodes a member of the Dbl family of oncogenes and controls cell shape changes during gastrulation in Drosophila, Genes Dev., 12, 274, 10.1101/gad.12.2.274
Hakeda-Suzuki, 2002, Rac function and regulation during Drosophila development, Nature, 416, 438, 10.1038/416438a
Han, 2002, Differential gene regulation by specific gain-of-function JNK1 proteins expressed in Swiss 3T3 fibroblasts, J. Biol. Chem., 277, 47167, 10.1074/jbc.M204270200
Harden, 2002, Signaling pathways directing the movement and fusion of epithelial sheets: lessons from dorsal closure in Drosophila, Differentiation, 70, 181, 10.1046/j.1432-0436.2002.700408.x
Hariharan, 2006, Regulation of imaginal disc growth by tumor-suppressor genes in Drosophila, Annu. Rev. Genet., 40, 335, 10.1146/annurev.genet.39.073003.100738
Hay, 1994, Expression of baculovirus P35 prevents cell death in Drosophila, Development, 120, 2121, 10.1242/dev.120.8.2121
Humbert, 2008, Control of tumourigenesis by the Scribble/Dlg/Lgl polarity module, Oncogene, 27, 6888, 10.1038/onc.2008.341
Igaki, 2009, Correcting developmental errors by apoptosis: lessons from Drosophila JNK signaling, Apoptosis, 14, 1021, 10.1007/s10495-009-0361-7
Igaki, 2006, Loss of cell polarity drives tumor growth and invasion through JNK activation in Drosophila, Curr. Biol., 16, 1139, 10.1016/j.cub.2006.04.042
Igaki, 2009, Intrinsic tumor suppression and epithelial maintenance by endocytic activation of Eiger/TNF signaling in Drosophila, Dev. Cell, 16, 458, 10.1016/j.devcel.2009.01.002
Jack, 1997, The genes raw and ribbon are required for proper shape of tubular epithelial tissues in Drosophila, Genetics, 147, 243, 10.1093/genetics/147.1.243
Johnston, 2009, Competitive interactions between cells: death, growth, and geography, Science, 324, 1679, 10.1126/science.1163862
Johnston, 1999, Increased activator protein-1 DNA binding and c-Jun NH2-terminal kinase activity in human breast tumors with acquired tamoxifen resistance, Clin. Cancer Res., 5, 251
Kanuka, 2005, Gain-of-function screen identifies a role of the Sec61alpha translocon in Drosophila postmitotic neurotoxicity, Biochim. Biophys. Acta, 1726, 225, 10.1016/j.bbagen.2005.06.020
Karim, 1998, Ectopic expression of activated Ras1 induces hyperplastic growth and increased cell death in Drosophila imaginal tissues, Development, 125, 1, 10.1242/dev.125.1.1
Ke, 2010, The c-Jun NH2-terminal kinase 2 plays a dominant role in human epidermal neoplasia, Cancer Res., 70, 3080, 10.1158/0008-5472.CAN-09-2923
Kennedy, 2003, Role of JNK in tumor development, Cell Cycle, 2, 199
Kerman, 2008, Ribbon modulates apical membrane during tube elongation through Crumbs and Moesin, Dev. Biol., 320, 278, 10.1016/j.ydbio.2008.05.541
Knoblich, 1994, Cyclin E controls S phase progression and its down-regulation during Drosophila embryogenesis is required for the arrest of cell proliferation, Cell, 77, 107, 10.1016/0092-8674(94)90239-9
Kockel, 1997, Jun in Drosophila development: redundant and nonredundant functions and regulation by two MAPK signal transduction pathways, Genes Dev., 11, 1748, 10.1101/gad.11.13.1748
Kurada, 1998, Ras promotes cell survival in Drosophila by downregulating hid expression, Cell, 95, 319, 10.1016/S0092-8674(00)81764-X
Laviolette, 2005, A genetic screen for suppressors of Drosophila NSF2 neuromuscular junction overgrowth, Genetics, 170, 779, 10.1534/genetics.104.035691
Lee, 2001, The role of Wingless signaling in establishing the anteroposterior and dorsoventral axes of the eye disc, Development, 128, 1519, 10.1242/dev.128.9.1519
Lee, 2001, Mosaic analysis with a repressible cell marker (MARCM) for Drosophila neural development, Trends Neurosci., 24, 251, 10.1016/S0166-2236(00)01791-4
Lee, 2000, Essential roles of Drosophila RhoA in the regulation of neuroblast proliferation and dendritic but not axonal morphogenesis, Neuron, 25, 307, 10.1016/S0896-6273(00)80896-X
Leong, 2009, scribble mutants promote aPKC and JNK-dependent epithelial neoplasia independently of Crumbs, BMC Biol., 7, 62, 10.1186/1741-7007-7-62
Liu, 2004, Nucleotide exchange factor ECT2 interacts with the polarity protein complex Par6/Par3/protein kinase Czeta (PKCzeta) and regulates PKCzeta activity, Mol. Cell Biol., 24, 6665, 10.1128/MCB.24.15.6665-6675.2004
Luo, 1994, Distinct morphogenetic functions of similar small GTPases: Drosophila Drac1 is involved in axonal outgrowth and myoblast fusion, Genes Dev., 8, 1787, 10.1101/gad.8.15.1787
Martin-Blanco, 1997, Regulation of cell differentiation by the Drosophila Jun kinase cascade, Curr. Opin. Genet. Dev., 7, 666, 10.1016/S0959-437X(97)80015-9
Massarwa, 2009, Apical secretion in epithelial tubes of the Drosophila embryo is directed by the Formin-family protein Diaphanous, Dev. Cell, 16, 877, 10.1016/j.devcel.2009.04.010
Mattila, 2005, Role of Jun N-terminal Kinase (JNK) signaling in the wound healing and regeneration of a Drosophila melanogaster wing imaginal disc, Int. J. Dev. Biol., 49, 391, 10.1387/ijdb.052006jm
Menendez, 2010, A tumor-suppressing mechanism in Drosophila involving cell competition and the Hippo pathway, Proc. Natl. Acad. Sci. USA, 107, 14651, 10.1073/pnas.1009376107
Miki, 1993, Oncogene ect2 is related to regulators of small GTP-binding proteins, Nature, 362, 462, 10.1038/362462a0
Mulinari, 2008, DRhoGEF2 and diaphanous regulate contractile force during segmental groove morphogenesis in the Drosophila embryo, Mol. Biol. Cell, 19, 1883, 10.1091/mbc.e07-12-1230
Neisch, 2010, Rho1 regulates apoptosis via activation of the JNK signaling pathway at the plasma membrane, J. Cell Biol., 189, 311, 10.1083/jcb.200912010
Nielsen, 2007, c-Jun NH2-terminal kinase 2 is required for Ras transformation independently of activator protein 1, Cancer Res., 67, 178, 10.1158/0008-5472.CAN-06-2801
O'Keefe, 2007, Egfr/Ras signaling regulates DE-cadherin/Shotgun localization to control vein morphogenesis in the Drosophila wing, Dev. Biol., 311, 25, 10.1016/j.ydbio.2007.08.003
O'Keefe, 2001, The pebble GTP exchange factor and the control of cytokinesis, Cell. Struct. Funct., 26, 619, 10.1247/csf.26.619
Olson, 1998, Signals from Ras and Rho GTPases interact to regulate expression of p21Waf1/Cip1, Nature, 394, 295, 10.1038/28425
Padash Barmchi, 2005, DRhoGEF2 regulates actin organization and contractility in the Drosophila blastoderm embryo, J. Cell Biol., 168, 575, 10.1083/jcb.200407124
Pagliarini, 2003, A genetic screen in Drosophila for metastatic behavior, Science, 302, 1227, 10.1126/science.1088474
Patch, 2009, A second-site noncomplementation screen for modifiers of Rho1 signaling during imaginal disc morphogenesis in Drosophila, PLoS One, 4, e7574, 10.1371/journal.pone.0007574
Prober, 2000, Ras1 promotes cellular growth in the Drosophila wing, Cell, 100, 435, 10.1016/S0092-8674(00)80679-0
Prober, 2002, Interactions between Ras1, dMyc, and dPI3K signaling in the developing Drosophila wing, Genes Dev., 16, 2286, 10.1101/gad.991102
Prokopenko, 1999, A putative exchange factor for Rho1 GTPase is required for initiation of cytokinesis in Drosophila, Genes Dev., 13, 2301, 10.1101/gad.13.17.2301
Rhiner, 2010, Flower forms an extracellular code that reveals the fitness of a cell to its neighbors in Drosophila, Dev. Cell, 18, 985, 10.1016/j.devcel.2010.05.010
Richardson, 1995, Ectopic cyclin E expression induces premature entry into S phase and disrupts pattern formation in the Drosophila eye imaginal disc, Development, 121, 3371, 10.1242/dev.121.10.3371
Richardson, 1993, A Drosophila G1-specific cyclin E homolog exhibits different modes of expression during embryogenesis, Development, 119, 673, 10.1242/dev.119.3.673
Ridley, 2004, Rho proteins and cancer, Breast Cancer Res. Treat., 84, 13, 10.1023/B:BREA.0000018423.47497.c6
Robinson, 2010, Crumbs regulates Salvador/Warts/Hippo signaling in Drosophila via the FERM-domain protein Expanded, Curr. Biol., 20, 582, 10.1016/j.cub.2010.03.019
Schiff, 2000, Oxidative stress and AP-1 activity in tamoxifen-resistant breast tumors in vivo, J. Natl. Cancer Inst., 92, 1926, 10.1093/jnci/92.23.1926
Settleman, 2001, Rac 'n Rho: the music that shapes a developing embryo, Dev. Cell, 1, 321, 10.1016/S1534-5807(01)00053-3
Shim, 2001, The Drosophila ribbon gene encodes a nuclear BTB domain protein that promotes epithelial migration and morphogenesis, Development, 128, 4923, 10.1242/dev.128.23.4923
Somers, 2003, A RhoGEF and Rho family GTPase-activating protein complex links the contractile ring to cortical microtubules at the onset of cytokinesis, Dev. Cell, 4, 29, 10.1016/S1534-5807(02)00402-1
Sorlie, 2001, Gene expression patterns of breast carcinomas distinguish tumor subclasses with clinical implications, Proc. Natl. Acad. Sci. USA, 98, 10869, 10.1073/pnas.191367098
Sorlie, 2003, Repeated observation of breast tumor subtypes in independent gene expression data sets, Proc. Natl. Acad. Sci. USA, 100, 8418, 10.1073/pnas.0932692100
Sotillos, 2004, DaPKC-dependent phosphorylation of Crumbs is required for epithelial cell polarity in Drosophila, J. Cell Biol., 166, 549, 10.1083/jcb.200311031
Sotiriou, 2003, Breast cancer classification and prognosis based on gene expression profiles from a population-based study, Proc. Natl. Acad. Sci. USA, 100, 10393, 10.1073/pnas.1732912100
Speck, 2003, Moesin functions antagonistically to the Rho pathway to maintain epithelial integrity, Nature, 421, 83, 10.1038/nature01295
Srivastava, 2007, Basement membrane remodeling is essential for Drosophila disc eversion and tumor invasion, Proc. Natl. Acad. Sci. USA, 104, 2721, 10.1073/pnas.0611666104
Stronach, 2005, Dissecting JNK signaling, one KKKinase at a time, Dev. Dyn., 232, 575, 10.1002/dvdy.20283
Sun, 2011, Regulation of Hippo signaling by Jun kinase signaling during compensatory cell proliferation and regeneration, and in neoplastic tumors, Dev. Biol., 350, 139, 10.1016/j.ydbio.2010.11.036
Titus, 2005, Rho proteins in cell migration and metastasis, Crit. Rev. Eukaryot. Gene Expr., 15, 103, 10.1615/CritRevEukaryotGeneExpr.v15.i2.20
Toba, 1999, The gene search system: a method for efficient detection and rapid molecular identification of genes in Drosophila melanogaster, Genetics, 151, 725, 10.1093/genetics/151.2.725
Uhlirova, 2006, JNK- and Fos-regulated Mmp1 expression cooperates with Ras to induce invasive tumors in Drosophila, EMBO J., 25, 5294, 10.1038/sj.emboj.7601401
Vidal, 2007, Differing Src signaling levels have distinct outcomes in Drosophila, Cancer Res., 67, 10278, 10.1158/0008-5472.CAN-07-1376
Wagner, 2009, Signal integration by JNK and p38 MAPK pathways in cancer development, Nat. Rev. Cancer, 9, 537, 10.1038/nrc2694
Warner, 2010, The Cdc42/Par6/aPKC polarity complex regulates apoptosis-induced compensatory proliferation in epithelia, Curr. Biol., 20, 677, 10.1016/j.cub.2010.03.025
Wasser, 2000, The EAST protein of Drosophila controls an expandable nuclear endoskeleton, Nat. Cell. Biol., 2, 268, 10.1038/35010535
Weber, 2000, Jun mediates Frizzled-induced R3/R4 cell fate distinction and planar polarity determination in the Drosophila eye, Development, 127, 3619, 10.1242/dev.127.16.3619
Weston, 2007, The JNK signal transduction pathway, Curr. Opin. Cell. Biol., 19, 142, 10.1016/j.ceb.2007.02.001
Whitmarsh, 2007, Role of mitogen-activated protein kinase kinase 4 in cancer, Oncogene, 26, 3172, 10.1038/sj.onc.1210410
Whyte, 2009, Key signalling nodes in mammary gland development and cancer: mitogen-activated protein kinase signalling in experimental models of breast cancer progression and in mammary gland development, Breast Cancer Res., 11, 209, 10.1186/bcr2361
Widmann, 2009, Dpp signaling promotes the cuboidal-to-columnar shape transition of Drosophila wing disc epithelia by regulating Rho1, J. Cell Sci., 122, 1362, 10.1242/jcs.044271
Woodhouse, 1998, Growth, metastasis, and invasiveness of Drosophila tumors caused by mutations in specific tumor suppressor genes, Dev. Genes Evol., 207, 542, 10.1007/s004270050145
Wu, 2010, Interaction between Ras(V12) and scribbled clones induces tumour growth and invasion, Nature, 463, 545, 10.1038/nature08702
Yan, 2009, Rho1 has multiple functions in Drosophila wing planar polarity, Dev. Biol., 333, 186, 10.1016/j.ydbio.2009.06.027
Zhang, 2006, Misexpression screen for genes altering the olfactory map in Drosophila, Genesis, 44, 189, 10.1002/dvg.20202
Zhang, 2007, Tumor necrosis factor receptor 1/c-Jun-NH2-kinase signaling promotes human neoplasia, Cancer Res., 67, 3827, 10.1158/0008-5472.CAN-06-4017