Proteomic and SAGE profiling of murine melanoma progression indicates the reduction of proteins responsible for ROS degradation

Authors

  • Gustavo A. de Souza,

    1. Centro de Terapia Celular, Centro Regional de Hemoterapia de Ribeirão Preto, Centro de Pesquisa, Inovação e Difusão (CEPID) – FAPESP, Brasil
    2. Centro de Quimica de Proteinas, Departamento de Biologia Celular, Molecular e Bioagentes Patogênicos, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP, Brasil
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  • Lyris M. F. Godoy,

    1. Centro de Terapia Celular, Centro Regional de Hemoterapia de Ribeirão Preto, Centro de Pesquisa, Inovação e Difusão (CEPID) – FAPESP, Brasil
    2. Centro de Quimica de Proteinas, Departamento de Biologia Celular, Molecular e Bioagentes Patogênicos, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP, Brasil
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  • Veronica R. Teixeira,

    1. Laboratorio de Oncologia Experimental LIM-24, Faculdade de Medicina, Universidade de São Paulo, São Paulo, SP, Brasil
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  • Andreia H. Otake,

    1. Laboratorio de Oncologia Experimental LIM-24, Faculdade de Medicina, Universidade de São Paulo, São Paulo, SP, Brasil
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  • Adão Sabino,

    1. Laboratorio Thomson de Espectrometria de Massa, Instituto de Química, Universidade de Campinas, Campinas, SP, Brasil
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  • José C. Rosa,

    1. Centro de Terapia Celular, Centro Regional de Hemoterapia de Ribeirão Preto, Centro de Pesquisa, Inovação e Difusão (CEPID) – FAPESP, Brasil
    2. Centro de Quimica de Proteinas, Departamento de Biologia Celular, Molecular e Bioagentes Patogênicos, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP, Brasil
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  • Anemari R. Dinarte,

    1. Centro de Terapia Celular, Centro Regional de Hemoterapia de Ribeirão Preto, Centro de Pesquisa, Inovação e Difusão (CEPID) – FAPESP, Brasil
    2. Laboratorio de Genética Molecular e BioInformática, Centro Regional de Hemoterapia de Ribeirão Preto, Ribeirão Preto, SP, Brasil
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  • Daniel G. Pinheiro,

    1. Centro de Terapia Celular, Centro Regional de Hemoterapia de Ribeirão Preto, Centro de Pesquisa, Inovação e Difusão (CEPID) – FAPESP, Brasil
    2. Laboratorio de Genética Molecular e BioInformática, Centro Regional de Hemoterapia de Ribeirão Preto, Ribeirão Preto, SP, Brasil
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  • Wilson A. Silva Jr.,

    1. Centro de Terapia Celular, Centro Regional de Hemoterapia de Ribeirão Preto, Centro de Pesquisa, Inovação e Difusão (CEPID) – FAPESP, Brasil
    2. Laboratorio de Genética Molecular e BioInformática, Centro Regional de Hemoterapia de Ribeirão Preto, Ribeirão Preto, SP, Brasil
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  • Marcos N. Eberlin,

    1. Laboratorio Thomson de Espectrometria de Massa, Instituto de Química, Universidade de Campinas, Campinas, SP, Brasil
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  • Roger Chammas Professor,

    1. Centro de Terapia Celular, Centro Regional de Hemoterapia de Ribeirão Preto, Centro de Pesquisa, Inovação e Difusão (CEPID) – FAPESP, Brasil
    2. Laboratorio de Oncologia Experimental LIM-24, Faculdade de Medicina, Universidade de São Paulo, São Paulo, SP, Brasil
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    • Additional corresponding author

  • Lewis J. Greene Professor

    Corresponding author
    1. Centro de Terapia Celular, Centro Regional de Hemoterapia de Ribeirão Preto, Centro de Pesquisa, Inovação e Difusão (CEPID) – FAPESP, Brasil
    2. Centro de Quimica de Proteinas, Departamento de Biologia Celular, Molecular e Bioagentes Patogênicos, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP, Brasil
    • Centro de Quimica de Proteinas, Fundacão Hemocentro de Ribeirão Preto, R.Ten. Catão Roxo 2501, 14051–140 Ribeirão Preto, SP, Brasil Fax: +55-16-2101-9366
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Abstract

Using 2-DE of total cell protein extracts, we compared soluble proteins from murine melanoma lines Tm1 and Tm5 with proteins from the nontumoral cell melan-a from which they were derived. Seventy-one of the 452 spots (average) detected with CBB were differentially accumulated, i.e., increased or decreased twofold. Forty-four spots were identified by PMF/MALDI-TOF, 15 with increased and 29 with decreased protein levels. SAGE showed that 17/34 (50%) of the differentially accumulated proteins, pI range 4–7, presented similar differences at the mRNA level. Major reductions in protein were observed in tumor cells of proteins that degrade reactive oxygen species (ROS). Decreases of ⋛ twofold in GST, superoxide dismutase, aldehyde dehydrogenase, thioredoxin, peroxiredoxin 2, and peroxiredoxin 6 protein were observed. SAGE indicated the reduction of other proteins involved in ROS degradation. As expected, the accumulation of exogenous peroxides was significantly higher in the tumor cells while the levels of glutathionylation were two times lower in the tumor cells compared to melan-a. The differential accumulation of proteins involved in oncogene/tumor suppressor pathways was observed. Melanoma cells can favor survival pathways activated by ROS by inhibiting p53 pathways and activation of Ras and c-myc pathways.

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