血管内皮生长因子165反义RNA治疗人脑胶质瘤的实验研究

目的 探讨应用血管内皮生长因子１６５（ＶＥＧＦ１６５）反义ＲＮＡ治疗人脑胶质瘤的可行性。方法 构建和鉴定反义ＶＥＧＦ１６５真核表达载体；将其转染入人脑胶质瘤细胞ＳＨＧ４４，检测其转染前、后的生物学性状；比较转染前、后ＳＨＧ４４细胞的裸鼠皮下致瘤性；分别应用免疫印迹、免疫组织化学、微血管计数、电镜和流式细胞仪检测上述改变。结果 成功构建反义ＶＥＧＦ１６５真核表达载体并在ＳＨＧ４４细胞获得表达，该细胞

Experimental research of gene therapy for human gliomas with vascular endothelial growth factor(165) antisense RNA]

OBJECTIVE: To investigate the feasibility of gene therapy for human gliomas with vascular endothelial growth factor(165) (VEGF(165)) antisense RNA. METHODS: The eukaryotic expression vector of antisense VEGF(165) was constructed and identified. Then the vector was transfected into human glioma cells (SHG44). The biological characteristics and tumorigenesis of SHG44 cells before and after transfection were inspected and compared. The changes were detected by Western blot, immunohistochemistry, micrangium counting, electron microscopy and flow cytometry. RESULTS: The eukaryotic expression vector pcDNA-AVEGF(165) was successfully constructed and transfected into SHG44 glioma cells. The characterstics of the cells were not influenced by the expression of antisense gene. The capability of tumorigenesis and angiogenesis of the transfected cells in nude mices was greatly reduced (tumour end volume, experiment group 212 mm(3); control group 7 897 mm(3); P < 0.01; Blood vessel counting: experiment group, 5.50; control group 11.22; P < 0.01 ). CONCLUSION: The angiogenesis and tumor growth of human gliomas are effectively inhibited by VEGF(165) antisense RNA. This experiment lays a foundation for solid tumor-specific gene therapy.