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Gastrin-Releasing Peptide Receptor Knockdown Induces Senescence in Glioblastoma Cells

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Abstract

Glioblastoma multiforme (GBM) is the most aggressive type of brain tumor, characterized by excessive cell proliferation, resistance to apoptosis, and invasiveness. Due to resistance to currently available treatment options, the prognosis for patients with GBM is very dismal. The activation of gastrin-releasing peptide receptors (GRPR) stimulates GBM cell proliferation, whereas GRPR antagonists induce antiproliferative effects in in vitro and in vivo experimental models of GBM. However, the role of GRPR in regulating other aspects of GBM cell function related to tumor progression remains poorly understood, and previous studies have not used RNA interference techniques as tools to examine GRPR function in GBM. Here, we found that stable GRPR knockdown by a lentiviral vector using a short hairpin interfering RNA sequence in human A172 GBM cells resulted in increased cell size and altered cell cycle dynamics consistent with cell senescence. These changes were accompanied by increases in the content of p53, p21, and p16, activation of epidermal growth factor receptors (EGFR), and a reduction in p38 content. These results increase our understanding of GRPR regulation of GBM cells and further support that GRPR may be a relevant therapeutic target in GBM.

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Acknowledgments

This research was supported by the National Council for Scientific and Technological Development (CNPq; grant numbers 484185/2012-8 and 303276/2013-4 to R. R), the National Institute for Translational Medicine (INCT-TM), the Children’s Cancer Institute (ICI), the South American Office for Anticancer Drug Development, and the HCPA institutional research fund (FIPE/HCPA).

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    Authors and Affiliations

    1. Cancer and Neurobiology Laboratory, Experimental Research Center, Clinical Hospital (CPE-HCPA), Federal University of Rio Grande do Sul, 90035-003, Porto Alegre, RS, Brazil

      Pâmela Rossi Menegotto, Patrícia Luciana da Costa Lopez, Bárbara Kunzler Souza, Caroline Brunetto de Farias, Gilberto Schwartsmann & Rafael Roesler

    2. Department of Pharmacology, Institute for Basic Health Sciences, Federal University of Rio Grande do Sul, Rua Sarmento Leite, 500 (ICBS, Campus Centro/UFRGS), 90050-170, Porto Alegre, RS, Brazil

      Pâmela Rossi Menegotto, Bárbara Kunzler Souza & Rafael Roesler

    3. Graduate Program in Gastroenterology and Hepatology, School of Medicine, Federal University of Rio Grande do Sul, Porto Alegre, RS, Brazil

      Patrícia Luciana da Costa Lopez & Eduardo Cremonese Filippi-Chiela

    4. Children’s Cancer Institute, 90420-140, Porto Alegre, RS, Brazil

      Caroline Brunetto de Farias

    5. Department of Biophysics, Laboratory of Cellular Plasticity and Signaling, Institute of Biosciences, and Center of Biotechnology, Federal University of Rio Grande do Sul, Porto Alegre, RS, Brazil

      Eduardo Cremonese Filippi-Chiela & Guido Lenz

    6. Laboratory of Genomic Medicine, Experimental Research Center, Clinical Hospital (CPE-HCPA), Federal University of Rio Grande do Sul, 90035-003, Porto Alegre, RS, Brazil

      Igor Araújo Vieira

    7. Department of Internal Medicine, Faculty of Medicine, Federal University of Rio Grande do Sul, 90035-003, Porto Alegre, RS, Brazil

      Gilberto Schwartsmann

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    1. Pâmela Rossi Menegotto
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    2. Patrícia Luciana da Costa Lopez
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    3. Bárbara Kunzler Souza
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    Correspondence to Rafael Roesler.

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    Pâmela Rossi Menegotto and Patrícia Luciana da Costa Lopez contributed equally to this work.

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    Menegotto, P.R., da Costa Lopez, P.L., Souza, B.K. et al. Gastrin-Releasing Peptide Receptor Knockdown Induces Senescence in Glioblastoma Cells. Mol Neurobiol 54, 888–894 (2017). https://doi.org/10.1007/s12035-016-9696-6

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    • DOI: https://doi.org/10.1007/s12035-016-9696-6

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