神经胶质瘤细胞中SWAP-70慢病毒载体的构建及表达

发布时间：2018-03-13 14:28

本文选题：SWAP-　切入点：神经胶质瘤　出处：《江苏医药》2016年21期 　论文类型：期刊论文

【摘要】：目的构建下调及过表达SWAP-70的慢病毒载体,并检测其在神经胶质瘤细胞中的表达效果。方法采用PCR扩增目的基因片段,构建SWAP-70慢病毒表达载体,与慢病毒转染系统共转染至293T细胞。收集病毒颗粒上清液,感染神经胶质瘤细胞。荧光显微镜下观察细胞中绿色荧光蛋白的表达情况,并采用Western blot法鉴定SWAP-70蛋白表达水平。结果成功构建下调及过表达SWAP-70的慢病毒载体,感染神经胶质瘤细胞的效率达90%左右。Western blot结果显示,下调SWAP-70的神经胶质瘤细胞中SWAP-70蛋白表达降低(P0.01),而过表达SWAP-70的神经胶质瘤细胞中内源性及外源性SWAP-70蛋白均有表达。结论成功建立下调及过表达SWAP-70的慢病毒载体,并有效转染神经胶质瘤细胞,为胶质瘤的基因治疗研究提供参考。
[Abstract]:Objective to construct a lentivirus vector for down-regulation and overexpression of SWAP-70 and to detect its expression in glioma cells. Methods SWAP-70 lentivirus expression vector was constructed by PCR amplification of the target gene fragment. Co-transfection with lentivirus transfection system to 293T cells. The supernatant of virus particles was collected to infect glioma cells. The expression of green fluorescent protein (GFP) in the cells was observed under fluorescence microscope. The expression level of SWAP-70 protein was identified by Western blot. Results Lentivirus vector was successfully constructed for down-regulation and overexpression of SWAP-70. The efficiency of infected glioma cells was about 90%. The down-regulation of SWAP-70 protein expression in glioma cells with SWAP-70 decreased the expression of SWAP-70 protein, while the expression of endogenous and exogenous SWAP-70 protein was found in glioma cells with overexpression of SWAP-70. Conclusion Lentivirus vector for down-regulation and overexpression of SWAP-70 is successfully established. And transfection of glioma cells to provide a reference for gene therapy of glioma.
【作者单位】：徐州医科大学研究生学院;徐州医科大学神经系统疾病研究所;徐州医科大学附属医院神经外科;
【基金】：国家自然科学基金青年基金(81402074)
【分类号】：R739.41

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