The Effect Of Angiostatin Gene Therapy And Chemotherapy On The Implanted Human Ovarian Carcinoma In Nude Mice

[Objectives]1. To investigate the effects of angiostatin gene therapy on subcutaneously implanted tumor induced by human ovarian epithelial carcinoma cell line SKOV3 in nude mouse.2. To observe the effects of angiostatin gene therapy combined with chemotherapy on intraperitoneal tumor and ascites fluid induced by human ovarian epithelial carcinoma cell line SKOV3 in nude mouse.[Materials and Methods]female nude mice, 4-5 weeks old, were used. The cell line of human ovarian epithelial carcinoma, named SKOV3, was cultured at 37℃ in 1640 medium, which supplemented with 10% calf serum. Tumors were established by injiection of about 5 ×10~6 and 1 ×10~7 tumor cells respectively into the back and abdominal cavity of nude mice.In the first experiment of angiostatin gene therapy used alone, the mice were classified randomly into three groups, and the growth of subcutaneous tumors were determined by measuring the long and short diameters everyday. Once the tumors reached 0.4cm in diameter, they were injected respectively with 100μl NS, empty plamid pcDNA3.0 and angiostatin plasmid. The transduction was mediated bycationic liposome DOTAP. In the second experiment of angiostatin gene therapy combined with chemotherapy, the mice were classified randomly into four groups after 7 days of the intraperitoneal injection of tumor cells, and injected respectively with empty plasmid pcDNA3.0 , angiostatin plasmid , cisplatin , angiostatin plasmid + cisplatin. For combinational treatment, reagents were delivered in a timed fashion, where angiostatin plasmid was injected first, followed by cisplatin 24h later.The tumor samples were prepared to be used in the examinations of the expression of angiostatin, VEGF with immunohistochemistry and Western Blot analysis, of MVD in the tumor with immunohistochemistry, and of cell apoptosis with TUNEL staining.Results were expressed as mean values ± standard deviation ( X ±s ). SNK test and x~2 test were used for evaluating statistical significance, where a value less than 0.05 (P<0.05) denote statistical significance. The software SPSS 11.5 was used in the statistical analysis. [Results]1. In the first experiment, tumor grew rapidly in the control group, reaching 1cm in size 18-21 days; whereas, in the angiostatin group, tumors were suppressed significantly after gene transfer , but they did not vanish and grow up slowly. Immunohistochemistry analysis and western blot analysis of tumor tissue revealed overexpression of angiostatin in situ. In addition, the expression of VEGF in the angiostatin group was a little higher than in the control group. AI in the angiostatin group was much higher than in the control group (P<0.01), concurrently MVD was lower (P<0.05).2. In the second experiment, there were 3 groups excluded control group could be found the inhibition of the tumor-growth and the ascites-formation. The action of the group combining angiostatin gene therapy with cisplatin was more significant than the other two groups. In this group, MVD was the lowest and AI was the highest. [Conclusion]...