Protein delivery of caspase-3 induces cell death in malignant C6 glioma, primary astrocytes and immortalized and primary brain capillary endothelial cells.
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 (beta-galactosidase) using lipid-mediated gene transfer (FuGENE6) or using the novel protein delivery reagent BioPORTER. beta-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 BioPORTER-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.
Zassler
Birgit
B
Laboratory of Psychiatry, University Clinic of Psychiatry, Innsbruck, Austria.
Blasig
Ingolf E
IE
Humpel
Christian
C
D000818:Animals; D001253:Astrocytes / Q000187:drug effects*; D001921:Brain / Q000098:blood supply*; D002196:Capillaries / Q000187:drug effects* / Q000473:pathology; D053148:Caspase 3; D020169:Caspases / Q000235:genetics* / Q000494:pharmacology*; D016923:Cell Death; D002461:Cell Line, Transformed; D045744:Cell Line, Tumor; D042783:Endothelial Cells / Q000187:drug effects* / Q000473:pathology; D018014:Gene Transfer Techniques; D005910:Glioma / Q000473:pathology / Q000503:physiopathology*; D006801:Humans; D020287:In Situ Nick-End Labeling; D051381:Rats; D013194:Staining and Labeling
C505407:CASP3 protein, human; C505410:Casp3 protein, rat; D053148:Caspase 3; D020169:Caspases