【摘要】 目的研究以腺相关病毒载体的反义血管内皮细胞生长因子(vascularendothelialgrowthfactor,VEGF)基因转染对视网膜母细胞瘤(retinoblastoma,RB)细胞中VEGF表达的影响,探讨基因治疗RB的新方法。方法将一定量的反义VEGF基因转染到RB细胞系HXORb44细胞,分别采用免疫组化图像分析和半定量RTPCR方法,检测转染前后HXORb44细胞中VEGF表达的变化。结果免疫组化显示VEGF蛋白在反义VEGF基因转染后的HXORb44细胞胞浆中的染色强度明显减弱,转染前的VEGF蛋白的灰度值为172.86±15.56,转染后的灰度值为243.41±18.62(灰度值与免疫染色强度成反比),差异有显著性意义(P=0.007)。半定量RTPCR表明VEGFmRNA在转染后的HXORb44细胞中的相对表达量为0.428±0.05,和转染前的表达量0.952±0.02相比,差异有显著性意义(P=0.000)。结论反义VEGF基因能显著抑制HXORb44细胞中VEGF的表达,为进一步的动物实验提供了实验依据。更多还原

【Abstract】 Objective To investigate the effect of antisense vascular endothelial growth factor (VEGF) gene transfection on the expression of VEGF in retinoblastoma cell by means of adeno-associated virus-mediated vector in vitro and to explore a new gene therapy strategy for retinoblastoma.Methods A small amounts of antisense VEGF genes were transfected into retinoblastoma cell line HXO-Rb 44,immunohistochemical image analysis and semi-quantitive RT-PCR techniques were used to evaluate the expression of VEGF in retinoblastoma cell line HXO-Rb 44.Results Immunohistochemical image analysis showed that the staining degree of VEGF protein decreased significantly after antisense VEGF genes transferred (the gray numerical value:17286±15.56 vs 243.41±18.62,P=0.007).Semi-quantitive RT-PCR measurements showed that the expression of VEGF mRNA decreased significantly after transfection (relative expression levels:0.952±0.02 vs 0.428±005,P=0000).Conclusion It is suggested that the antisense VEGF genes have an inhibitory effect on the expression of VEGF in HXO-Rb 44 cell in vitro and the results provided the experimental evidence for the further treatment of retinoblastoma.更多还原