OBJECTIVE: To discuss the feasibility of gene therapy of human glioma by antiangiogenesis method. METHODS: Angiostatin K(1-3) cDNA with secretive signal was inserted into the polylinker sites of eukaryotic expression vector pcDNA3 to construct pcDNA-SAK(1-3). The vector was transfected into human SHG44 glioma cells by lipofectamine and the positive clone was screened by G418. The biological characteristics of glioma cells were examined by electronmicroscope and flow cytometry. The activity of angiostatin K(1-3) protein expressed by SHG44 cells was examined by the bovine micrangium endotheliocyte inhibition assay and immunofluorescence assay. When SHG44 cells were implanted into the strata subcutaneum of nude mice, tumor necrosis and micrangium were calculated immunohistochemically and electronmicroscopically for determining their characteristics and validity in gene therapy of human glioma by antiangiogenesis method. RESULTS: The eukaryotic expression vector pcDNA-SAK (1-3) was successfully constructed and transfected into glioma cells. The cells expressed angiostatin K(1-3) protein, and their tumorigenesis and angiogenesis in nude mice were greatly reduced. CONCLUSION: Angiostatin K(1-3) gene is feasible to treat human glioma. This experiment lays a foundation for gene therapy of the other solid tumors by antiangiogenesis method.
OBJECTIVE: To discuss the feasibility of gene therapy of humanglioma by antiangiogenesis method. METHODS: Angiostatin K(1-3) cDNA with secretive signal was inserted into the polylinker sites of eukaryotic expression vector pcDNA3 to construct pcDNA-SAK(1-3). The vector was transfected into human SHG44 glioma cells by lipofectamine and the positive clone was screened by G418. The biological characteristics of glioma cells were examined by electronmicroscope and flow cytometry. The activity of angiostatin K(1-3) protein expressed by SHG44 cells was examined by the bovine micrangium endotheliocyte inhibition assay and immunofluorescence assay. When SHG44 cells were implanted into the strata subcutaneum of nude mice, tumor necrosis and micrangium were calculated immunohistochemically and electronmicroscopically for determining their characteristics and validity in gene therapy of humanglioma by antiangiogenesis method. RESULTS: The eukaryotic expression vector pcDNA-SAK (1-3) was successfully constructed and transfected into glioma cells. The cells expressed angiostatin K(1-3) protein, and their tumorigenesis and angiogenesis in nude mice were greatly reduced. CONCLUSION: Angiostatin K(1-3) gene is feasible to treat humanglioma. This experiment lays a foundation for gene therapy of the other solid tumors by antiangiogenesis method.
Authors: Oszkar Szentirmai; Cheryl H Baker; Szofia S Bullain; Ning Lin; Masaya Takahashi; Judah Folkman; Richard C Mulligan; Bob S Carter Journal: J Neurosurg Date: 2008-05 Impact factor: 5.115