Adenovirus-mediated transfer of bax with caspase-8 controlled by myelin basic protein promoter exerts an enhanced cytotoxic effect in gliomas.
The transduction of Bax protein, which is up-regulated in radiation- and chemotherapy-induced apoptosis, augments the cytotoxicity of radiotherapy and chemotherapy for cancers. The cytotoxicity of Bax overexpression is caused primarily by mitochondrial dysfunction, which is also involved in the apoptosis triggered by caspase-8. In this study, we transduced the Bax gene in combination with caspase-8 gene to evaluate whether or not this approach induces effective cytotoxicity in glioma cells. In terms of cancer gene therapy, it is critically important to induce cytotoxic genes in a cancer-specific manner. Therefore, we used the myelin basic protein promoter to drive cytotoxic genes. The expression level controlled by the myelin basic protein promoter was relatively low in gliomas. In U251 and U-373 MG glioma cells, adenovirus-mediated transduction of the Bax gene combined with caspase-8 gene induced enhanced apoptosis and cell death as determined by morphological analysis and assay for dead cells, hypodiploid cells, and DNA fragmentation (terminal deoxynucleotidyltransferase-mediated deoxyuridine triphosphate nick end labeling method). This therapeutic modality would be useful to induce a specific and enhanced cytotoxic effect for gliomas.
Shinoura
N
N
Department of Molecular Biotherapy Research, Cancer Chemotherapy Center, Japanese Foundation for Cancer Research, Tokyo.
Saito
K
K
Yoshida
Y
Y
Hashimoto
M
M
Asai
A
A
Kirino
T
T
Hamada
H
H
D000256:Adenoviridae / Q000235:genetics*; D001932:Brain Neoplasms / Q000628:therapy*; D053181:Caspase 8; D053453:Caspase 9; D020169:Caspases / Q000235:genetics* / Q000378:metabolism; D016923:Cell Death; D002469:Cell Separation; D053938:DNA Fragmentation; D018076:DNA, Complementary / Q000378:metabolism; D005434:Flow Cytometry; D018014:Gene Transfer Techniques; D005910:Glioma / Q000628:therapy*; D006801:Humans; D015151:Immunoblotting; D007763:Lac Operon; D008854:Microscopy, Electron; D008928:Mitochondria / Q000378:metabolism; D004676:Myelin Basic Protein / Q000235:genetics*; D011401:Promoter Regions, Genetic; D011518:Proto-Oncogene Proteins / Q000235:genetics* / Q000378:metabolism; D019253:Proto-Oncogene Proteins c-bcl-2; D014161:Transduction, Genetic; D014407:Tumor Cells, Cultured; D051028:bcl-2-Associated X Protein
C494826:BAX protein, human; D018076:DNA, Complementary; D004676:Myelin Basic Protein; D011518:Proto-Oncogene Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D051028:bcl-2-Associated X Protein; C505530:CASP8 protein, human; C506433:CASP9 protein, human; D053181:Caspase 8; D053453:Caspase 9; D020169:Caspases