ABSTRACT
OBJECTIVE@#To investigate the in vitro and in vivo anticancer efficacy of the immunotoxin DTAT and DTATEGF against globlastoma multiforme.@*METHODS@#The in vitro cytotoxicity of DTAT and DTATEGF was measured using MTT assay. In vivo studies were performed in which 18 nude mice were randomly divided into 3 groups and the glioma xenograft intracranial mouse model was constructed with U87-luc cell line of human glioma. Then 1 μg of DTAT, or DTATEGF, or a control protein Bickel3 was delivered intracranially by convection-enhanced delivery (CED) via an osmotic minipump. The brain tumor fluorescence signal intensity was investigated by bioluminescent imaging (BLI). Microvessel density (MVD) was measured by immunchistochemistry SABC method in each group.@*RESULTS@#In vitro DTAT and DTATEGF were found highly potent against U87-luc cell line, with IC(50) <0.01 nmol/L and IC(50)<1 nmol/L, respectively. In vivo BLI monitoring of the control group showed progressively increasing luminescence, while in the two treatment groups, luminescence was reduced on day 8, and increased slowly (P<0.05). The MVD of DTAT (31.6±5.2)/mm(2) and DTATEGF (25.1±6.5)/mm(2) groups had significant difference with that of the control group (51.3±7.4) /mm(2) (P<0.01).@*CONCLUSION@#Both DTAT and DTATEGF have potential in clinical application against globlastoma multiforme because of their ability to target the tumor cells and neovasculature simultaneously.
Subject(s)
Animals , Humans , Mice , Angiogenesis Inhibitors , Pharmacology , Brain Neoplasms , Drug Therapy , Cell Line, Tumor , Glioblastoma , Drug Therapy , Glioma , Immunotoxins , Pharmacology , Mice, Nude , Xenograft Model Antitumor AssaysABSTRACT
OBJECTIVE@#To investigate the in vitro and in vivo anticancer efficacy of the immunotoxin DTATEGF against human NSCLC brain metastatic tumor PC9-BrM3 cell line.@*METHODS@#The effect of the immunotoxin DTATEGF was tested for its ability to inhibit the proliferation of PC9-BrM3 cells in vitro by MTT assay. The cell cycle and the apoptosis of cells with 1 pmol/L DTATEGF were examined by flow cytometry. In vivo, 2 μg of DTATEGF or control Bickel3 was given intratumor to nude mice with established PC9-BrM3 xenografts on their hips, and tumor volumes were measured and tumor samples were investigated by immunchistochemistry SABC method. The microvessel density (MVD) was measured in each group.@*RESULTS@#In vitro, DTATEGF killed PC9-BrM3 cells and showed an IC50 of 1 pmol/L. The apoptotic rate in the 1 pmol/L DTATEGF group was (64.0±0.5)% , significantly higher than that in the control group (1.5±0.4)% (P<0.01). The cell cycle was obviously inhibited by DTATEGF in a dose-dependent manner. The percentage of cells treated with 1 pmol/L DTATEGF in SubG0/G1 phase was (32.0±1.5)%, significantly higher than that in the control group (2.0±0.4)% (P<0.01). In vivo, DTATEGF significantly inhibited the growth of PC9-BrM3 hip tumors (P<0.05). The MVD of the DTATEGF group was (15.6±4.6)/mm2, significantly lower than that of the control group (31.2±5.4)/mm2 (P<0.01).@*CONCLUSION@#DTATEGF inhibits the growth of the PC9-BrM3 cell line and induces its apoptosis. It is highly efficacious against human metastatic NSCLC brain tumor and against neovascularization.