Objective To investigate the mRNA-based anticancer gene transfer in MSCs-mediated cytotox-icity of glioma cells. Methods TRAIL mRNA and PTEN mRNA were synthesized in vitro. Immunoblotting assay was used to detect the expression of TRAIL and PTEN in the transfected MSCs.Transwell co-culture was perform to detect the migration ability of MSCs after gene transfection. The bioluminescence,live/dead staining and real time cell analyzer were used to analyze the viability of DBTRG cells. Results Compared with non-transfected MSCs, an enhanced migration rate was observed in MSCs with two kind of mRNA transfection.TRAIL-and PTEN-mRNA-induced cytotoxicity in DBTRG glioma cell was correlated with the ratio of the conditioned medium of the transfect-ed MSCs. A synergistic action was observed in TRAIL and PTEN in the transwell co-culture model. Conclusion The present study reveals the effect of synthesized mRNA-based gene transfer on mesenchymal stem cell-mediated cytotoxicity of glioma cells(DBTRG).%目的 探讨利用体外合成抑癌基因TRAIL-、PTEN-mRNA编辑的间充质干细胞(MSCs)对DBTRG细胞杀伤作用.方法 利用体外反转录方法合成TRAIL-、PTEN-mRNA,转染MSC后用western blotting方法检测TARIL及PTEN蛋白的表达情况,利用transwell共培养方法观察mRNA编辑的MSCs对DBTRG的迁移趋向性作用,利用荧光显微镜和活体成像仪检测在间接共培养条件下TRAIL-、PTEN-mRNA编辑的MSCs对DBTRG-LUC细胞的杀伤作用.结果 体外合成的mRNA均能在MSCs中正确表达TRAIL、PTEN蛋白;与对照组对比两种mRNA编辑的MSCs对DBTRG细胞的趋向性均明显增强;TRAIL-、PTEN-mRNA编辑的MSCs培养上清(CM)均能显著抑制DBTRG细胞生长,并且TRAIL和PTEN共表达组存在着协同作用.结论 利用体外合成抑癌基因mRNA的方法研究MSCs介导的双基因靶向对神经胶质瘤的杀伤作用,为今后临床利用人工合成的多基因mRNA靶向治疗肿瘤提供全新思路.
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