Intra-tumoral administration of naked plasmid DNA encoding mouse endostatin inhibits renal carcinoma growth.
Endostatin is a C-terminal fragment of collagen XVIII and has potent anti-angiogenic and anti-tumor activity. Mouse endostatin-coding sequences were obtained using PCR and linked to the signal sequence of influenzavirus hemagglutinin. The signal-sequence endostatin fragment was subcloned into plasmid vectors under the transcriptional control of cytomegalovirus promoter. Murine renal carcinoma (Renca) cells transfected with endostatin-coding plasmid are shown to secrete full-length endostatin. Endostatin-secreting Renca cells demonstrate slower growth in vivo compared to empty vector-transfected cells, but their in vitro growth is unaffected. Anti-angiogenic activity of secreted endostatin was confirmed in a Matrigel angiogenesis assay in vivo. We report growth inhibition of Renca tumors resulting from intra-tumoral delivery of plasmid vector encoding secretable endostatin. Elevated local concentrations of endostatin resulted from multiple intra-tumoral injections of endotoxin-purified plasmid DNA. Local endostatin levels were high enough to obtain growth arrest of Renca tumors.
Department of Molecular Biology, Centre of Oncology, Maria Sklodowska-Curie Memorial Institute, 44-101 Gliwice, Poland.
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D000818:Animals; D015153:Blotting, Western; D002292:Carcinoma, Renal Cell / Q000473:pathology / Q000517:prevention & control*; D003094:Collagen / Q000235:genetics*; D043170:Collagen Type XVIII; D004247:DNA / Q000008:administration & dosage*; D017931:DNA Primers / Q000737:chemistry; D043169:Endostatins; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D015316:Genetic Therapy; D007279:Injections, Subcutaneous; D007680:Kidney Neoplasms / Q000473:pathology / Q000517:prevention & control*; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D009368:Neoplasm Transplantation; D009389:Neovascularization, Pathologic; D010446:Peptide Fragments / Q000235:genetics*; D010957:Plasmids / Q000235:genetics*; D016133:Polymerase Chain Reaction; D014162:Transfection; D014407:Tumor Cells, Cultured
D043170:Collagen Type XVIII; D017931:DNA Primers; D043169:Endostatins; D010446:Peptide Fragments; D003094:Collagen; D004247:DNA