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Concomitant treatment with pertussis toxin plus temozolomide increases the survival of rats bearing intracerebral RG2 glioma

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Abstract

Purpose

Glioblastoma multiforme is the most frequent primary brain tumor, it has poor prognosis, and it remains refractory to current treatment. The success of temozolomide (TMZ) appears to be limited by the occurrence of chemoresistance. Recently, we report the use of pertussis toxin as adjuvant immunotherapy in a C6 glioma model; showing a decrease in tumoral size, it induced selective cell death in Treg cells, and it elicited less infiltration of tumoral macrophages. Here, we evaluated the cytotoxic effect of pertussis toxin in combination with TMZ for glioma treatment, both in vitro and in vivo RG2 glioma model.

Methods

We determined cell viability, cell cycle, apoptosis, and autophagy on treated RG2 cells through flow cytometry, immunofluorescence, and Western blot assays. Twenty-eight rats were divided in four groups (n = 7) for each treatment. After intracranial implantation of RG2 cells, animals were treated with TMZ (10 mg/Kg/200 μl of apple juice), PTx (2 μg/200 μl of saline solution), and TMZ + PTx. Animals without treatment were considered as control.

Results

We found an induction of apoptosis in around 20 % of RG2 cells, in both single treatments and in their combination. Also, we determined the presence of autophagy vesicles, without any modifications in the cell cycle in the TMZ – PTx-treated groups. The survival analyses showed an increase due to individual treatments; while in the group treated with the combination TMZ − PTx, this effect was enhanced.

Conclusion

We show that the concomitant use of pertussis toxin plus TMZ could represent an advantage to improve the glioma treatment.

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Acknowledgments

Animals were kindly donated by Bioinvert Mexico City. This work was supported by the National Council of Science and Technology of Mexico (CONACyT, Grant 180851) and Instituto de Ciencia y Tecnología del Distrito Federal (PICSA 10-143).

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Authors and Affiliations

  1. Neuroimmunology and Neuro-Oncology Unit, Instituto Nacional de Neurología y Neurocirugía (INNN), Insurgentes Sur 3877, 14269, Mexico City, Mexico

    Roxana Magaña-Maldonado, Norma Y. Hernández-Pedro, Julio Sotelo & Benjamín Pineda

  2. Immunology Departament, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México (UNAM), Mexico City, Mexico

    Roxana Magaña-Maldonado & Karen Manoutcharian

  3. Excitatory Amino Acids Laboratory, Instituto Nacional de Neurología y Neurocirugía (INNN), Mexico City, Mexico

    Edgar Rangel-López

  4. Neurochemistry Unit, Instituto Nacional de Neurología y Neurocirugía (INNN), Mexico City, Mexico

    Verónica Pérez-De la Cruz

  5. Animal Housing Unit, Instituto Nacional de Neurología y Neurocirugía (INNN), Mexico City, Mexico

    César Rodríguez-Balderas

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  1. Roxana Magaña-Maldonado
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  2. Karen Manoutcharian
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Correspondence to Benjamín Pineda.

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Magaña-Maldonado, R., Manoutcharian, K., Hernández-Pedro, N.Y. et al. Concomitant treatment with pertussis toxin plus temozolomide increases the survival of rats bearing intracerebral RG2 glioma. J Cancer Res Clin Oncol 140, 291–301 (2014). https://doi.org/10.1007/s00432-013-1565-3

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  • DOI: https://doi.org/10.1007/s00432-013-1565-3

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