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BUB1 and BUBR1 inhibition decreases proliferation and colony formation, and enhances radiation sensitivity in pediatric glioblastoma cells

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Abstract

Purpose

Glioblastoma (GBM) is a very aggressive and lethal brain tumor with poor prognosis. Despite new treatment strategies, patients’ median survival is still lower than 1 year in most cases. The expression of the BUB gene family has demonstrated to be altered in a variety of solid tumors, pointing to a role as putative therapeutic target. The purpose of this study was to determine BUB1, BUB3, and BUBR1 gene expression profiles in glioblastoma and to analyze the effects of BUB1 and BUBR1 inhibition combined or not with Temozolomide and radiation in the pediatric SF188 GBM cell line.

Methods

For gene expression analysis, 8 cell lines and 18 tumor samples were used. The effect of BUB1 and BUBR1 inhibition was evaluated using siRNA. Apoptosis, cell proliferation, cell cycle kinetics, micronuclei formation, and clonogenic capacity were analyzed after BUB1 and BUBR1 inhibition. Additionally, combinatorial effects of gene inhibition and radiation or Temozolomide (TMZ) treatment were evaluated through proliferation and clonogenic capacity assays.

Results

We report the upregulation of BUB1 and BUBR1 expression and the downregulation of BUB3 in GBM samples and cell lines when compared to white matter samples (p < 0.05). Decreased cell proliferation and colony formation after BUB1 and BUBR1 inhibition were observed, along with increased micronuclei formation. Combinations with TMZ also caused cell cycle arrest and increased apoptosis. Moreover, our results demonstrate that BUB1 and BUBR1 inhibition sensitized SF188 cells to γ-irradiation as shown by decreased growth and abrogation of colony formation capacity.

Conclusion

BUB1 and BUBR1 inhibition decreases proliferation and shows radiosensitizing effects on pediatric GBM cells, which could improve treatment strategies for this devastating tumor. Collectively, these findings highlight the potentials of BUB1 and BUBR1 as putative therapeutic targets for glioblastoma treatment.

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Acknowledgments

We are grateful to BSc Veridiana Kiill Suazo for technical assistance. We also thank the Fundação de Apoio a Pesquisa do Estado de São Paulo (FAPESP; 2009/50118-2 and 2009/09305-3) and the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) for financial support.

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Authors and Affiliations

  1. Department of Genetics, University of São Paulo, Av. Bandeirantes, 3900, Bairro Monte Alegre, 14048-900, Ribeirão Preto, Sao Paulo, Brazil

    Andressa Gois Morales, Julia Alejandra Pezuk & Jaqueline Carvalho de Oliveira

  2. Division of Pediatric Oncology, Department of Pediatrics, Hospital das Clínicas da Faculdade de Medicina de Ribeirão Preto–USP, Laboratório de Pediatria, Bloco G. Av. Bandeirantes, 3900, Bairro Monte Alegre, 14048-900, Ribeirão Preto, Sao Paulo, Brazil

    María Sol Brassesco, Rosane Gomes de Paula Queiroz, Carlos Alberto Scrideli & Luiz Gonzaga Tone

  3. Department of Surgery and Anatomy, Faculty of Medicine of Ribeirao Preto, University of São Paulo, Av. Bandeirantes, 3900, Bairro Monte Alegre, 14048-900, Ribeirão Preto, Sao Paulo, Brazil

    Hélio Rubens Machado & Carlos Gilberto Carlotti Jr

  4. Department of Pathology, Faculty of Medicine of Ribeirao Preto, University of São Paulo, Av. Bandeirantes, 3900, Bairro Monte Alegre, 14048-900, Ribeirão Preto, Sao Paulo, Brazil

    Luciano Neder

  5. Clinics Department, Faculty of Medicine of Ribeirão Preto, University of São Paulo, Av. Bandeirantes, 3900, Bairro Monte Alegre, 14048-900, Ribeirão Preto, Sao Paulo, Brazil

    Harley Francisco de Oliveira

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  1. Andressa Gois Morales
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  2. Julia Alejandra Pezuk
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  3. María Sol Brassesco
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  4. Jaqueline Carvalho de Oliveira
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  5. Rosane Gomes de Paula Queiroz
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  7. Carlos Gilberto Carlotti Jr
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  8. Luciano Neder
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  9. Harley Francisco de Oliveira
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  11. Luiz Gonzaga Tone
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Corresponding author

Correspondence to María Sol Brassesco.

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AG Morales and JA Pezuk contributed equally to this paper.

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Morales, A.G., Pezuk, J.A., Brassesco, M.S. et al. BUB1 and BUBR1 inhibition decreases proliferation and colony formation, and enhances radiation sensitivity in pediatric glioblastoma cells. Childs Nerv Syst 29, 2241–2248 (2013). https://doi.org/10.1007/s00381-013-2175-8

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