血管内皮生长因子_(165)反义RNA治疗人脑胶质瘤的实验研究

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

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 ,该细胞生物学性状不受外源基因表达影响 ,其在裸鼠皮下致瘤性和血管生成能力明显下降。肿瘤终体积 :实验组 2 12mm3,对照组 7897mm3(P <0 .0 1) ;血管计数 :实验组 5 .5 0 ,对照组11 2 2 (P <0 .0 1)。结论 VEGF165反义RNA能有效抑制人脑胶质瘤血管生成和肿瘤生长 。

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 antiense RNA. This experiment lays a foundation for solid tumor specific gene therapy.