脂质体介导B7基因在C6胶质瘤细胞中表达的实验研究
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Q78 R730


Expression of B7 gene in C6 Mediated with Liposome
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    摘要:

    目的 通过脂质体介导基因转移方法,将含B7基因的重组逆转录病毒表达质粒导入C6细胞中。使之能有效的表达B7分子,探索胶质瘤免疫基因治疗的新方法。方法 用脂质体将pLXSN—B7重组表达质粒导入Wistar大鼠胶质瘤细胞系C6中,经G418筛选出阳性克隆。通过PCR、狭缝杂交及B7单克隆抗体免疫荧光方法检测B7基因在C6细胞中的整合、转录及表达情况。结果 PCR可从B7基因转导的C6细胞(C6pLXSN-B7)基因组中扩增出0.8 kb的B7基因片断;以B7为探针狭缝杂交显示C6pLXSN—B7细胞中有B7基因的转录;以抗-B7单克隆抗体进行的免疫荧光检测显示C6pLXSN—B7细胞表面有B7分子的表达,而对照组肿瘤细胞(C6pLXSN、C6)则无B7分子的表达。结论 脂质体介导B7基因转移方法可将B7基因完整整合到C6细胞基因组中并使之能稳定、有效的表达。

    Abstract:

    Objective To explore a new immunological genetic therapy for glioma with the liposome-mediated transgenic method.Methods We introduced the pLXSN-expressed recombinated plasmid into the glioma cell line C6 of Wistar rat,then we pick up the positive clone by G418 screening,finally,got the expression of B7 gene in C6 cell line with PCR,RNA-slot hybridization.Results With PCR,we obtained 0.8Kb's segment of B7 gene from the genome of the B7-transdued C6 cell (C6pLXSN-B7); with RNA-slot hybridization using B7 as the probe,we found the transcription of B7 gene in the C6pLXSN-B7 cells;with anti-B7 the expression of B7 molecule on the surface of C6pLXSN-B7 cells,on the contrary,there was no expression of B7 in the glioma cells of the control group (C6pLXSN,C6).Conclusion The liposome-mediated B7 transgenic method can lead to the entire integration of B7 gene into the genome of C6 cells and the resulting stable and effective expression of B7 gene.

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李永宁 幸兵 任祖渊 苏长保 王任直 马文斌.脂质体介导B7基因在C6胶质瘤细胞中表达的实验研究[J].中国比较医学杂志,2001,(1):35~38.

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  • 最后修改日期:2000-11-19
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