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Effects of tumor biobank storage on polysome stability
Lupinacci, Fernanda C S; Ferreira, Elisa N; Roffe, Martin; Bellato, Hermano M; Carraro, Dirce Maria; Hajj, Glaucia N M.
  • Lupinacci, Fernanda C S; A.C.Camargo Cancer Center. International Research Center. São Paulo. BR
  • Ferreira, Elisa N; A.C.Camargo Cancer Center. International Research Center. São Paulo. BR
  • Roffe, Martin; A.C.Camargo Cancer Center. International Research Center. São Paulo. BR
  • Bellato, Hermano M; A.C.Camargo Cancer Center. International Research Center. São Paulo. BR
  • Carraro, Dirce Maria; A.C.Camargo Cancer Center. International Research Center. São Paulo. BR
  • Hajj, Glaucia N M; A.C.Camargo Cancer Center. International Research Center. São Paulo. BR
Appl. cancer res ; 39: 1-5, 2019. ilus, tab
Artigo em Inglês | LILACS, Inca | ID: biblio-1015230
Biblioteca responsável: BR30.1
ABSTRACT

Background:

Human biological material has become an important resource for biomedical research. Tumor Biobanks are facilities that collect, store and distribute samples of tumor and normal tissue for further use in basic and translational cancer research. mRNA-translation has been demonstrated to modulate protein levels and is considered a fundamental post-transcriptional mechanism of gene expression regulation. Thus, determining translation efficiencies of individual mRNAs in human tumors may add another layer of information that contributes to the understanding of tumorigenic pathways. To analyze the RNAs actively engaged in translation, RNAs associated with ribosomes (polysomes) are isolated, identified and compared to total RNA. However, the application of this technique in human tumors depends on the stability of the polysomal structure under Biobank storage conditions that usually consists of ultra-low temperature. Since the effect of freezing on the stability of the polysomal structure in stored tumor samples is not known, it is essential to evaluate this factor in the frozen samples, validating the use of biobank samples in studies of translational efficiency.

Methods:

Xenograft tumors were divided in two parts, half was subject to immediate processing, and half was frozen for posterior analysis. Both parts were subject to polysomal separation, RNA extraction and identification through RNAseq.

Results:

It was possible to successfully extract and identify total and polysomal RNA from both fresh and frozen tumoral tissue. The quantification of the polysome profile indicated no difference in the translational efficiency estimated in fresh versus frozen tissue. Gene expression data from the fresh versus frozen tissues were compared and the correlation between the polysome associated fresh x frozen (R = 0,89) and total fresh x frozen (0,90) mRNAs was calculated. No difference was identified between the two conditions.

Conclusions:

We demonstrated that tissue freezing does not affect the polysomal structure, consequently validating the viability of the use of biobank stored tissue for polysome associated RNA analysis (AU)
Assuntos


Texto completo: Disponível Coleções: Bases de dados internacionais Temas: Geral / Tipos de Câncer / Outros Tipos Base de dados: LILACS / PrevCan Assunto principal: Polirribossomos / RNA / Expressão Gênica / Regulação da Expressão Gênica / Neoplasias Tipo de estudo: Prognostic_studies Limite: Humanos Idioma: Inglês Revista: Appl. cancer res Assunto da revista: Neoplasias Ano de publicação: 2019 Tipo de documento: Artigo País de afiliação: Brasil Instituição/País de afiliação: A.C.Camargo Cancer Center/BR

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Texto completo: Disponível Coleções: Bases de dados internacionais Temas: Geral / Tipos de Câncer / Outros Tipos Base de dados: LILACS / PrevCan Assunto principal: Polirribossomos / RNA / Expressão Gênica / Regulação da Expressão Gênica / Neoplasias Tipo de estudo: Prognostic_studies Limite: Humanos Idioma: Inglês Revista: Appl. cancer res Assunto da revista: Neoplasias Ano de publicação: 2019 Tipo de documento: Artigo País de afiliação: Brasil Instituição/País de afiliação: A.C.Camargo Cancer Center/BR