Adam17 mediates OSCC development in an orthotopic murine model

Background: ADAM17 is one of the main sheddases of the cells and it is responsible for the cleavage and the release of ectodomains of important signaling molecules, such as EGFR ligands. Despite the known crosstalk between ADAM17 and EGFR, which has been considered a promising targeted therapy in oral squamous cell carcinoma (OSCC), the role of ADAM17 in OSCC development is not clear. Method: In this study the effect of overexpressing ADAM17 in cell migration, viability, adhesion and proliferation was comprehensively appraised in vitro. In addition, the tumor size, tumor proliferative activity, tumor collagenase activity and MS-based proteomics of tumor tissues have been evaluated by injecting tumorigenic squamous carcinoma cells (SCC-9) overexpressing ADAM17 in immunodeficient mice. Results: The proteomic analysis has effectively identified a total of 2,194 proteins in control and tumor tissues. Among these, 110 proteins have been down-regulated and 90 have been up-regulated in tumor tissues. Biological network analysis has uncovered that overexpression of ADAM17 regulates Erk pathway in OSCC and further indicates proteins regulated by the overexpression of ADAM17 in the respective pathway. These results are also supported by the evidences of higher viability, migration, adhesion and proliferation in SCC-9 or A431 cells in vitro along with the increase of tumor size and proliferative activity and higher tissue collagenase activity as an outcome of ADAM17 overexpression. Conclusion: These findings contribute to understand the role of ADAM17 in oral cancer development and as a potential therapeutic target in oral cancer. In addition, our study also provides the basis for the development of novel and refined OSCC-targeting approaches. © 2014 Simabuco et al.; licensee BioMed Central Ltd.ADAM17 is one of the main sheddases of the cells and it is responsible for the cleavage and the release of ectodomains of important signaling molecules, such as EGFR ligands. Despite the known crosstalk between ADAM17 and EGFR, which has been considered a1311324FAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOCNPQ - CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO2009/54067-3; 2010/19278-0; 2010/15675-5470567/2009-0; 470549/2011-4; 301702/2011-

Integrated Proteomics Identified Up-regulated Focal Adhesion-mediated Proteins In Human Squamous Cell Carcinoma In An Orthotopic Murine Model

Understanding the molecular mechanisms of oral carcinogenesis will yield important advances in diagnostics, prognostics, effective treatment, and outcome of oral cancer. Hence, in this study we have investigated the proteomic and peptidomic profiles by combining an orthotopic murine model of oral squamous cell carcinoma (OSCC), mass spectrometry-based proteomics and biological network analysis. Our results indicated the up-regulation of proteins involved in actin cytoskeleton organization and cell-cell junction assembly events and their expression was validated in human OSCC tissues. In addition, the functional relevance of talin-1 in OSCC adhesion, migration and invasion was demonstrated. Taken together, this study identified specific processes deregulated in oral cancer and provided novel refined OSCC-targeting molecules. © 2014 Granato et al.95Siegel, R., Naishadham, D., Jemal, A., Cancer statistics, 2013 (2013) CA Cancer J Clin, 63, pp. 11-30Wong, D.T.W., Todd, R., Tsuji, T., Donoff, R.B., Molecular biology of human oral cancer (1996) Critical Reviews in Oral Biology and Medicine, 7 (4), pp. 319-328Warnakulasuriya, S., Global epidemiology of oral and oropharyngeal cancer (2009) Oral Oncol, 45, pp. 309-316Choi, S., Myers, J.N., Molecular pathogenesis of oral squamous cell carcinoma: Implications for therapy (2008) J Dent Res, 87, pp. 14-32Panis, C., Pizzatti, L., Herrera, A.C., Cecchini, R., Abdelhay, E., Putative circulating markers of the early and advanced stages of breast cancer identified by high-resolution label-free proteomics (2013) Cancer Lett, 330, pp. 57-66Fujita, Y., Nakanishi, T., Miyamoto, Y., Hiramatsu, M., Mabuchi, H., Proteomics-based identification of autoantibody against heat shock protein 70 as a diagnostic marker in esophageal squamous cell carcinoma (2008) Cancer Lett, 263, pp. 280-290Mognetti, B., Di Carlo, F., Berta, G.N., Animal models in oral cancer research (2006) Oral Oncol, 42, pp. 448-460Hwang, Y.S., Zhang, X., Park, K.K., Chung, W.Y., An orthotopic and osteolytic model with a newly established oral squamous cell carcinoma cell line (2012) Arch Oral Biol, 58, pp. 218-225Boukamp, P., Petrussevska, R.T., Breitkreutz, D., Hornung, J., Markham, A., Fusenig, N.E., Normal keratinization in a spontaneously immortalized aneuploid human keratinocyte cell line (1988) Journal of Cell Biology, 106 (3), pp. 761-771Agostini, M., Almeida, L.Y., Bastos, D.C., Ortega, R.M., Moreira, F.S., The fatty acid synthase inhibitor orlistat reduces the growth and metastasis of orthotopic tongue oral squamous cell carcinomas (2014) Mol Cancer Ther, 13, pp. 585-595Villen, J., Gygi, S.P., The SCX/IMAC enrichment approach for global phosphorylation analysis by mass spectrometry (2008) Nat Protoc, 3, pp. 1630-1638Paes Leme, A.F., Sherman, N.E., Smalley, D.M., Sizukusa, L.O., Oliveira, A.K., Hemorrhagic activity of HF3, a snake venom metalloproteinase: Insights from the proteomic analysis of mouse skin and blood plasma (2012) J Proteome Res, 11, pp. 279-291Aragao, A.Z., Nogueira, M.L., Granato, D.C., Simabuco, F.M., Honorato, R.V., Identification of novel interaction between ADAM17 (a disintegrin and metalloprotease 17) and thioredoxin-1 (2012) J Biol Chem, 287, pp. 43071-43082(2009) R: A Language and Environment for Statistical Computing. 2.11.1 Ed., , Team RDC Vienna: R Foundation for Statistical ComputingZhang, B., VerBerkmoes, N.C., Langston, M.A., Uberbacher, E., Hettich, R.L., Samatova, N.F., Detecting differential and correlated protein expression in label-free shotgun proteomics (2006) Journal of Proteome Research, 5 (11), pp. 2909-2918. , DOI 10.1021/pr0600273Cox, J., Mann, M., MaxQuant enables high peptide identification rates, individualized p.P.B.-range mass accuracies and proteome-wide protein quantification (2008) Nat Biotechnol, 26, pp. 1367-1372Kanehisa, M., Araki, M., Goto, S., Hattori, M., Hirakawa, M., Itoh, M., Katayama, T., Yamanishi, Y., KEGG for linking genomes to life and the environment (2008) Nucleic Acids Research, 36 (SUPPL. 1), pp. D480-D484. , DOI 10.1093/nar/gkm882Dennis Jr., G., Sherman, B.T., Hosack, D.A., Yang, J., Gao, W., DAVID: Database for Annotation, Visualization, and Integrated Discovery (2003) Genome Biol, 4, pp. P3Wang, Z., Jiang, L., Huang, C., Li, Z., Chen, L., Comparative proteomics approach to screening of potential diagnostic and therapeutic targets for oral squamous cell carcinoma (2008) Mol Cell Proteomics, 7, pp. 1639-1650Wetting, H.L., Hadler-Olsen, E., Magnussen, S., Rikardsen, O., Steigen, S.E., S100A4 expression in xenograft tumors of human carcinoma cell lines is induced by the tumor microenvironment (2011) Am J Pathol, 178, pp. 2389-2396Myers, J.N., Holsinger, F.C., Jasser, S.A., Bekele, B.N., Fidler, I.J., An orthotopic nude mouse model of oral tongue squamous cell carcinoma (2002) Clinical Cancer Research, 8 (1), pp. 293-298Heath, C.H., Deep, N.L., Sweeny, L., Zinn, K.R., Rosenthal, E.L., Use of panitumumab-IRDye800 to image microscopic head and neck cancer in an orthotopic surgical model (2012) Ann Surg Oncol, 19, pp. 3879-3887Wang, Y., McNiven, M.A., Invasive matrix degradation at focal adhesions occurs via protease recruitment by a FAK-p130Cas complex (2012) J Cell Biol, 196, pp. 375-385Lai, M.T., Hua, C.H., Tsai, M.H., Wan, L., Lin, Y.J., Talin-1 overexpression defines high risk for aggressive oral squamous cell carcinoma and promotes cancer metastasis (2011) J Pathol, 224, pp. 367-376Drees, F., Pokutta, S., Yamada, S., Nelson, W.J., Weis, W.I., Alpha-catenin is a molecular switch that binds E-cadherin-beta-catenin and regulates actin-filament assembly (2005) Cell, 123 (5), pp. 903-915. , DOI 10.1016/j.cell.2005.09.021, PII S009286740500975XKiema, T., Lad, Y., Jiang, P., Oxley, C.L., Baldassarre, M., Wegener, K.L., Campbell, I.D., Calderwood, D.A., The molecular basis of filamin binding to integrins and competition with talin (2006) Molecular Cell, 21 (3), pp. 337-347. , DOI 10.1016/j.molcel.2006.01.011, PII S1097276506000311Yue, J., Lu, H., Liu, J., Berwick, M., Shen, Z., Filamin-A as a marker and target for DNA damage based cancer therapy (2012) DNA Repair (Amst), 11, pp. 192-200Warburg, O., On the origin of cancer cells (1956) Science, 123, pp. 309-314Jones, R.G., Thompson, C.B., Tumor suppressors and cell metabolism: A recipe for cancer growth (2009) Genes Dev, 23, pp. 537-548Hanahan, D., Weinberg, R.A., Hallmarks of cancer: The next generation (2011) Cell, 144, pp. 646-674Israel, M., Schwartz, L., The metabolic advantage of tumor cells (2011) Mol Cancer, 10, p. 70Cantor, J.R., Sabatini, D.M., Cancer cell metabolism: One hallmark, many faces (2012) Cancer Discov, 2, pp. 881-898Shen, Y., Tolic, N., Xie, F., Zhao, R., Purvine, S.O., Effectiveness of CID, HCD, and ETD with FT MS/MS for degradomic- peptidomic analysis: Comparison of peptide identification methods (2011) J Proteome Res, 10, pp. 3929-3943Assinder, S.J., Stanton, J.A., Prasad, P.D., Transgelin: An actin-binding protein and tumour suppressor (2009) Int J Biochem Cell Biol, 41, pp. 482-486Lynch, C.D., Lazar, A.M., Iskratsch, T., Zhang, X., Sheetz, M.P., Endoplasmic spreading requires coalescence of vimentin intermediate filaments at force-bearing adhesions (2013) Mol Biol Cell, 24, pp. 21-30Gelman, J.S., Sironi, J., Castro, L.M., Ferro, E.S., Fricker, L.D., Peptidomic analysis of human cell lines (2011) J Proteome Res, 10, pp. 1583-1592Findeisen, P., Neumaier, M., Functional protease profiling for diagnosis of malignant disease (2012) Proteomics Clin Appl, 6, pp. 60-78Doucet, A., Butler, G.S., Rodriguez, D., Prudova, A., Overall, C.M., Metadegradomics: Toward in vivo quantitative degradomics of proteolytic post-translational modifications of the cancer proteome (2008) Mol Cell Proteomics, 7, pp. 1925-195

Soil bulk density and biomass partitioning of Brachiaria decumbens in a silvopastoral system Densidade do solo e partição de biomassa de Brachiaria decumbens em um sistema silvopastoril

Shade in silvopastoral systems improves the thermal comfort of animals, but it may also affect the pasture productivity and can contribute to soil compaction in the shaded areas due to the increase in the number of animals looking for comfort. The effect of grazing at various distances from tree rows (under the tree canopy, at 6 and at 12 m away from the trees) on the soil bulk density and on the aerial and root biomass of Brachiaria decumbens was evaluated in both the dry and the rainy seasons. The study was carried out on an Orthic Ferralsol in a randomized block design with two replications. Tree rows were composed of Eucalyptus grandis and Acacia mangium species, and the paddocks were submitted to a rotational stocking management, using Holstein (Bos taurus) × Zebu (Bos indicus) heifers. The shade intensity in the pasture decreased with an increasing distance from the tree row. Soil bulk density did not vary with the distance from the tree row, but varied seasonally, being greater in the rainy season (1.47 g cm-3) than in the dry season (1.28 g cm-3). Green forage and root mass, expressed as dry matter, were lower under the tree canopy and were greater in the rainy season. There were decreases of 22.3 and 41.4% in the aerial and root biomasses, respectively, in the tree rows. The greatest shoot/root ratio for B. decumbens under moderate and intensive shading indicates a modification in the forage biomass allocation pattern that favours the aerial development in detriment of the root system.<br>O sombreamento em sistemas silvipastoris concorre para o conforto térmico dos animais; no entanto pode afetar a produção do pasto e contribuir para a compactação do solo, pelo aumento da concentração de animais nas áreas sombreadas. Avaliou-se o efeito da distância do renque de árvores (sob a copa das árvores, 6 e 12 m de distancia das árvores) na densidade do solo e na biomassa aérea e de raízes de Brachiaria decumbens, nas épocas seca e chuvosa. O estudo foi conduzido num Latossolo Vermelho-Amarelo no delineamento em blocos casualizados, com duas repetições. A faixa de árvores foi composta pelas espécies Eucalyptus grandis e Acacia mangium, e os piquetes foram manejados com novilhas Holandês (Bos taurus) × Zebu (Bos indicus) , sob lotação rotativa. A intensidade de sombreamento foi decrescente com o distanciamento do renque de árvores. A densidade do solo não variou com a distância do renque de árvores, mas sim com a época do ano, tendo sido maior na época chuvosa (1,47 g dm-3) do que na seca (1,28 g dm-3). As massas secas de forragem verde e de raízes foram menores sob a copa das árvores e maiores na época chuvosa do que na seca. Na faixa arborizada houve reduções de 22,3 e 41,4% na biomassa aérea e de raízes, respectivamente. A maior relação parte aérea/raiz da B. decumbens à sombra expressa uma modificação no padrão de alocação de biomassa na forrageira, que prioriza a formação da parte aérea, em detrimento do sistema radicular