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。
Objective To investigate the effect of antiscnsc 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 antiscnsc VEGF genes were transfected into retinoblastonm cell line HXO-Rb44, immunohistochemical image analysis and scmi-quantitive RT-PCR techniques were used to evaluate the expression of VEGF in retinoblastoma cell line HXO-Rb44. Results Immunohistochemical image analysis showed that the staining degree of VFGF protein decreased significantly after antiscnsc 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 antiscnse VEGF genes have an inhibitory effect on the expression of VFGF in HXO-Rb44 cell in vitro and the results provided the experimental evidence for the further treatment of retinoblastoma.
出处
《眼科新进展》
CAS
2005年第4期338-340,共3页
Recent Advances in Ophthalmology