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目的探讨反义生存素(Survivin)脂质体复合物对脑胶质瘤细胞增殖和凋亡效应的影响及机制,为脑胶质瘤Surviviu基因靶向治疗提供理论依据。方法用脂质体介导Survivin反义寡核苷酸转染脑胶质瘤细胞系U251细胞;Western印迹试验检测Survivin表达水平变化;噻唑蓝 (MTT)法测量细胞生长情况;流式细胞仪检测细胞凋亡率及线粒体膜电位变化;电镜倒置显微镜观察细胞形态学变化;应用激光共聚焦显微镜免疫荧光分析法评价脂质体反义复合物对Survivin 蛋白的影响。结果反义Survivin脂质体复合物能有效下调Survivin表达水平,肿瘤细胞凋亡率随转染时间延长而逐渐递增,并出现凋亡形态学变化。免疫荧光分析中标记有绿色荧光染色的Sur- vivin蛋白分子在未转染的细胞浆中清晰可见,并呈斑点状分布;而在转染细胞中几乎没有发现。结论靶向抑制Survivin基因在脑胶质瘤未来治疗中有良好的应用前景。
Objective To investigate the effect and mechanism of antisense survivin liposome complex on proliferation and apoptosis of glioma cells and to provide a theoretical basis for the targeted therapy of glioma Surviviu gene. Methods Liposome-mediated Survivin antisense oligonucleotide was transfected into glioma cell line U251, Western blotting was used to detect the expression of Survivin, cell growth was measured by MTT assay, flow cytometry Cell apoptosis rate and mitochondrial membrane potential. The morphological changes of the cells were observed under inverted microscope. The effect of antisense liposome complex on Survivin protein was evaluated by confocal laser scanning microscopy. Results The antisense Survivin liposome complex could effectively down-regulate the expression of Survivin. The apoptosis rate of tumor cells gradually increased with the prolongation of transfection time, and the morphological changes of apoptosis appeared. Immunofluorescence analysis of green fluorescent staining of Survivin protein molecules in the untransfected cytoplasm was clearly visible and spotted distribution; and found in the transfected cells were scarcely found. Conclusion Targeted inhibition of Survivin gene in glioma future treatment has a good prospect.