Delivery of hypoxia and glioma dual-specific suicide gene using dexamethasone conjugated polyethylenimine for glioblastoma-specific gene therapy.
Gene therapy has been considered a promising approach for glioblastoma therapy. To avoid side effects and increase the specificity of gene expression, gene expression should be tightly regulated. In this study, glioma and hypoxia dual-specific plasmids (pEpo-NI2-SV-Luc and pEpo-NI2-SV-HSVtk) were developed by combining the erythropoietin (Epo) enhancer and nestin intron 2 (NI2). In the in vitro studies, pEpo-NI2-SV-Luc showed higher gene expression under hypoxia than normoxia in a glioblastoma-specific manner. The MTT and caspase assays demonstrated that pEpo-NI2-SV-HSVtk specifically induced caspase activity and cell death in hypoxic glioblastoma cells. For in vivo evaluation, subcutaneous and intracranial glioblastoma models were established. Dexamethasone-conjugated-polyethylenimine (PEI-Dexa) was used as a gene carrier, since PEI-Dexa efficiently delivers plasmid to glioblastoma cells and also has an antitumor effect due to the effect of dexamethasone. In the in vivo study in the subcutaneous and intracranial glioblastoma models, the tumor size was reduced more effectively in the pEpo-NI2-SV-HSVtk group than in the control and pSV-HSVtk groups. In addition, higher levels of HSVtk gene expression and TUNEL-positive cells were observed in the pEpo-NI2-SV-HSVtk group compared with the control and pSV-HSVtk groups, suggesting that pEpo-NI2-SV-HSVtk increased the therapeutic efficacy in hypoxic glioblastoma. Therefore, pEpo-NI2-SV-HSVtk/PEI-Dexa complex may be useful for glioblastoma-specific gene therapy.
Department of Bioengineering, College of Engineering, Hanyang University , Seoul 133-791, Republic of Korea.
Hyun Ah
HA
Kim
Jin Hyeong
JH
Park
Na
N
Yi
Minhyung
M
Lee
D000818:Animals; D017209:Apoptosis; D015153:Blotting, Western; D049109:Cell Proliferation; D002478:Cells, Cultured; D003907:Dexamethasone / Q000008:administration & dosage* / Q000493:pharmacokinetics; D004337:Drug Carriers; D024202:Electrophoretic Mobility Shift Assay; D004742:Enhancer Elements, Genetic / Q000235:genetics; D000068817:Epoetin Alfa; D004921:Erythropoietin / Q000235:genetics; D005434:Flow Cytometry; D018014:Gene Transfer Techniques; D041722:Genes, Transgenic, Suicide; D005909:Glioblastoma / Q000235:genetics / Q000473:pathology / Q000628:therapy*; D057809:HEK293 Cells; D006801:Humans; D000860:Hypoxia; D007124:Immunoenzyme Techniques; D007438:Introns / Q000235:genetics; D008156:Luciferases / Q000378:metabolism; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D064231:Nestin / Q000235:genetics; D010957:Plasmids / Q000008:administration & dosage; D011094:Polyethyleneimine / Q000737:chemistry*; D011401:Promoter Regions, Genetic / Q000235:genetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000235:genetics; D013937:Thymidine Kinase / Q000235:genetics*; D014018:Tissue Distribution; D014162:Transfection; D023041:Xenograft Model Antitumor Assays
D004337:Drug Carriers; D064231:Nestin; D011994:Recombinant Proteins; D004921:Erythropoietin; D000068817:Epoetin Alfa; D003907:Dexamethasone; D011094:Polyethyleneimine; D008156:Luciferases; D013937:Thymidine Kinase
gene regulation; gene therapy; glioblastoma; hypoxia; suicide gene