【摘要】 目的探讨TTA1、Tat、聚乙二醇(PEG)修饰的明胶硅氧烷纳米粒子(GS NPs)结合siRNA-EGFR转染C6细胞,抑制其表皮生长因子受体(EGFR)表达的影响。方法根据鼠EGFR的mRNA序列设计合成siRNA-EGFR以及阴性对照siRNA-Scr(无意义序列)。TTA1-Tat-PEG-GS NPs通过静电作用结合siRNA。实验分4组:Control组、TTA1-Tat-PEG-GS NPs/siRNA-Scr组、TTA1-Tat-PEG-GS NPs/siRNA-EGFR组及X-tremeGene/siRNA-EGFR组。除Control组外,其余三组均在体外对C6细胞进行转染。Real-time PCR、Western blot分别检测细胞EGFR mRNA及蛋白表达情况;CCK-8法、流式细胞仪分别检测细胞增殖和凋亡情况。结果与Control组及TTA1-Tat-PEG-GS NPs/siRNA-Scr组比较,TTA1-Tat-PEG-GS NPs/siRNA-EGFR组在体外细胞水平上能有效抑制C6细胞EGFR mRNA及蛋白的表达(P<0.01),同时抑制C6细胞的增殖(P<0.05或P<0.01)并促进其凋亡(P<0.01)。此外,X-tremeGene/siRNA-EGFR组和TTA1-Tat-PEG-GS NPs/siRNA-EGFR组在下调C6细胞EGFR水平、抑制细胞增殖及促进细胞凋亡方面差异无统计学意义(P>0.05)。结论 TTA1-Tat-PEG-GS NPs可能成为一种新型的基因治疗载体,为胶质瘤基因治疗的进一步研究提供理论依据。更多还原

【Abstract】 Objective To investigate the property of TTAl-Tat-PEG modified gelatin-siloxane nanoparticles(GS NPs)combined with siRNA-EGFR in transfection of C6 cells and inhibiting its EGFR gene expression.Methods The siRNA-EGFR and negative control sequences(siRNA-Scr) were designed and synthesized according to the EGFR mRNA sequence of rats.TTAl-Tat-PEG-GS NPs bound to siRNA via electrostatic interaction.The composites of experiment included control group,TTAl-Tat-PEG-GS NPs/siRNA-Scr group,TTAl-Tat-PEG-GS NPs/siRNA-EGFR group and X-tremeGene/siRNA-EGFR group.C6 cells were transfected in all groups except for those in the control group.Realtime PCR and Western blot were used to detect EGFR mRNA and protein levels,respectively.The proliferation and apoptosis were detected with CCK-8 and flowcytometry,respectively.Results Compared with control group and TTAlTat-PEG-GS NPs/siRNA-Scr group,TTA1-Tat-PEG-GS NPs/siRNA-EGFR group could inhibit the expression of EGFR mRNA and protein of C6 cells(P < 0.01),restrain the proliferation of C6 cells(P < 0.05 or P < 0.01) and promote its apoptosis(P < 0.01).In addition,there were no statistically significant differences between X-tremeGene/siRNA-EGFR group and TTA1-Tat-PEG-GS NPs/siRNA-EGFR group in decreasing the EGFR levels of C6 cells,inhibiting the cell proliferation and promoting the cell apoptosis(P > 0.05).Conclusion TTAl-Tat-PEG-GS NPs may be a novel gene carrier and provide theoretical foundation for the further research of glioma gene therapy.更多还原