title: Investigation of a plasmid containing a novel immunotoxin VEGF165-PE38 gene for antiangiogenic therapy in a malignant glioma model.
abstract: Inhibition of tumor neovascularization has profound effects on the growth of solid tumors. Our previous studies have shown the effect of VEGF165-PE38 recombinant immunotoxin on proliferation and apoptosis in human umbilical vein endothelial cells in vitro. In this study, we explored the direct inhibition of angiogenesis in chick chorioallantoic membrane and antiangiogenic therapy in a malignant glioma model. HEK293 cells were transfected with the pVEGF165PE38-IRES2-EGFP plasmid. ELISA was used to confirm the expression of VEGF165-PE38 in the transfected cells. These cells released 1396 + or - 131.9 pg VEGF165-PE38/1x10(4) cells/48 h into the culture medium and the supernatant was capable of inhibiting the growth of capillary-like structures in chick chorioallantoic membrane assay. In a murine malignant glioma model, plasmid was directly administered via multiple local intratumoral delivery. After day 16 the tumor volume in mice treated with pVEGF165PE38-IRES2-EGFP was significantly lower than that in mice in the control groups. Immunohistochemistry studies showed that the treated group had decreased expression of CD31. Quantitative analysis of microvessel density in the treated group was 1.99 + or - 0.69/0.74 mm(2), and was significantly lower than that in the control groups (9.33 + or - 1.99/0.74 mm(2), 8.09 + or - 1.39/0.74 mm(2) and 8.49 + or - 1.69/0.74 mm(2)). Immunohistochemistry analysis indicated that immunotoxin VEGF165-PE38 was distributed in the treated group in malignant glioma tissue. Our findings provide evidence that the in vivo production of VEGF165-PE38 through gene therapy using a eukaryotic expression plasmid had potential antiangiogenic activity in malignant glioma in vivo.
authors.affiliation: Department of Neurosurgery, Zhujiang Hospital, Southern Medical University, Guangzhou, China.
authors.forename: Chang-chen
authors.initials: CC
authors.lastname: Hu
authors.forename: Hong-ming
authors.initials: HM
authors.lastname: Ji
authors.forename: Sheng-li
authors.initials: SL
authors.lastname: Chen
authors.forename: Han-wei
authors.initials: HW
authors.lastname: Zhang
authors.forename: Bin-quan
authors.initials: BQ
authors.lastname: Wang
authors.forename: Li-yuan
authors.initials: LY
authors.lastname: Zhou
authors.forename: Zi-ping
authors.initials: ZP
authors.lastname: Zhang
authors.forename: Xin-lin
authors.initials: XL
authors.lastname: Sun
authors.forename: Zhen-zhou
authors.initials: ZZ
authors.lastname: Chen
authors.forename: Ying-qian
authors.initials: YQ
authors.lastname: Cai
authors.forename: Ling-sha
authors.initials: LS
authors.lastname: Qin
authors.forename: Li
authors.initials: L
authors.lastname: Lu
authors.forename: Xiao-dan
authors.initials: XD
authors.lastname: Jiang
authors.forename: Ru-xiang
authors.initials: RX
authors.lastname: Xu
authors.forename: Yi-quan
authors.initials: YQ
authors.lastname: Ke
mesh_terms: D036002:ADP Ribose Transferases / Q000627:therapeutic use; D020533:Angiogenesis Inhibitors / Q000627:therapeutic use*; D000818:Animals; D001427:Bacterial Toxins / Q000627:therapeutic use; D045744:Cell Line, Tumor; D005098:Exotoxins / Q000627:therapeutic use; D005240:Feasibility Studies; D015316:Genetic Therapy; D005910:Glioma / Q000098:blood supply / Q000628:therapy*; D006801:Humans; D000922:Immunotoxins / Q000627:therapeutic use*; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D010957:Plasmids; D011549:Pseudomonas / Q000378:metabolism; D014162:Transfection; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics*; D037521:Virulence Factors / Q000627:therapeutic use; D023041:Xenograft Model Antitumor Assays; D000097668:Pseudomonas aeruginosa Exotoxin A
chemical_list: D036002:ADP Ribose Transferases; D020533:Angiogenesis Inhibitors; D001427:Bacterial Toxins; D005098:Exotoxins; D000922:Immunotoxins; D042461:Vascular Endothelial Growth Factor A; D037521:Virulence Factors; D000097668:Pseudomonas aeruginosa Exotoxin A; C467484:VEGFA protein, human