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Protein delivery of caspase-3 induces cell death in malignant C6 glioma, primary astrocytes and immortalized and primary brain capillary endothelial cells

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Journal of Neuro-Oncology Aims and scope Submit manuscript

Abstract

Most brain tumors consist of transformed glia cells and are highly vascularized by capillary endothelial cells. The aim of the present study therefore was to deliver pro-apoptotic caspase-3 into malignant C6 glioma and immortalized rBCEC4 brain endothelial cells to induce cell death. Both cell lines were transfected with a reporter protein (β-galactosidase) using lipid-mediated gene transfer (FuGENE6TM) or using the novel protein delivery reagent BioPORTERTM. β-Galactosidase protein was successfully delivered into both cells, the protein expression peaked around day 2 and was transient. Delivery of caspase-3 induced TUNEL-positive cell death of both cell types. As a control, caspase-3 was also delivered to non-neoplastic primary astrocytes and endothelial cells and induced cell death. In conclusion BioPORTERTM-protein delivery of pro-apoptotic molecules may provide a potent tool to cause death of the cells in brain tumors, however, this method is limited due to its toxicity to non-malignant cells.

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Abbreviations

rBCEC4:

rat brain capillary endothelial cell line 4

C6:

cells from a rat glial tumor

PBS:

phosphate buffered saline

TBS:

tris buffered saline

FCS:

fetal calf serum

BSA:

bovine serum albumin

DMEM:

Dulbecco’s Modified Eagle’s Medium

pEF-ßGAL:

plasmid with the elongation factor alpha1 encoding ß-galactosidase

pEF(-):

control plasmid without functional gene

VEGF:

vascular endothelial growth factor

ECGF:

endothelial cell growth factor

ABC:

avidin biotin complex

DAB:

diaminobenzidine

X-GAL:

5-bromochloro-3-indolyl-β-d-galactopyranoside

TUNEL:

DNA Nick-End Labeling

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Authors and Affiliations

  1. Laboratory of Psychiatry, University Clinic of Psychiatry, Innsbruck, Austria

    Birgit Zassler & Christian Humpel

  2. Institute of Molecular Pharmacology, Berlin, Germany

    Ingolf E. Blasig

  3. Department of Psychiatry, Laboratory of Psychiatry, University Clinic of Psychiatry, Anichstr.35, A-6020, Innsbruck, Austria

    Christian Humpel

Authors
  1. Birgit Zassler
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  2. Ingolf E. Blasig
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  3. Christian Humpel
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Correspondence to Christian Humpel.

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Zassler, B., Blasig, I.E. & Humpel, C. Protein delivery of caspase-3 induces cell death in malignant C6 glioma, primary astrocytes and immortalized and primary brain capillary endothelial cells. J Neurooncol 71, 127–134 (2005). https://doi.org/10.1007/s11060-004-1364-4

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