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[摘要]
目的:研究EGF-R介导的靶向性非病毒载体GE7系统治疗人脑胶质瘤的体内外效应。方法:组建GE7基因转移系统,分别与β-gal报道基因、p21WAF-1/CIP1基因组成复合物,体外转导U251MG细胞,分析GE7系统导入效率;MTS法观察细胞生长曲线;FACS分析细胞周期变化。建立裸鼠皮下荷瘤模型,分别经瘤内和尾静脉注射导入DNA载体复合物,观察肿瘤生长抑制率。结果:GE7系统体外导入率最高达70%,细胞生长曲线呈低平型,细胞周期阻滞于G1期,平均凋亡指数为25.2%;瘤内注射和尾静脉注射抑瘤率分别为56.5%和60.2%。结论:GE7系统具有较高效率和靶向性,经肿瘤局部和全身途径导入p21WAF-1/CIP1基因,均可诱导凋亡,抑制肿瘤生长。
[Key word]
[Abstract]
Objective:To investigate in vitro and in vivo effects on growth of human gliomas mediated by the EGF-R targeting non-viral vector GE7 system.Methods:To construct GE7 system, U251MG cell line was transfected in vitro with β-galactosidase gene and p21WAF-1/CIP1 gene mediated by GE7 system. By means of X-gal staining, MTS, FACS, the transferring rate of exogenous gene and the apoptosis rate of tumor cells were examined. The animal model was established by implanting U251 cells subcutaneously in nude mouse. The U251MG glioma-bearing animals were injected with GE7/DNA/HA20 complexes subcutaneously and introvenously. One week later animals were killed, observed the average weight of each group and tumor inhibitory rate in different groups.Results:The highest transferring rate of GE7 system in vitro was 70%,and the expression of p21WAF-1/CIP1 gene could induce the apoptosis of the glioma cell and inhibit its growth. The average apoptosis rate was 25.2%. The subcutaneous and introvenous injection of GE7 system had the same therapeutic effect on human gliomas with the tumor inhibitory rate of 56.5% and 60.2%. Conclusion:This study provides a more effective way to improve the effeciency of glioma gene therapy.
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[基金项目]
本课题受国家“863”高科技项目(102-12-02-05)资助