TTA1―Tat―PEG―GS NPs结合siRNA―EGFR转染C6细胞抑制EGFR基因表达研究.docVIP

TTA1―Tat―PEG―GS NPs结合siRNA―EGFR转染C6细胞抑制EGFR基因表达研究 　　[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 0.05）。 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。 　　[关键词] 胶质瘤；纳米粒子；基因载体；RNA干扰 　　[中图分类号] R739.41 [文献标识码] A [文章编号] 1673-7210（2016）02（c）-0004-05 　　Research of TTA1-Tat-PEG-GS NPs combined with siRNA-EGFR in transfection of C6 cells and inhibiting its EGFR gene expression 　　HU Yang1 ZHANG Long1 LI Wang1 WEI Feng2 FENG Wei2 TIAN Xinhua2 　　1.Medical College， Xiamen University， Fujian Province， Xiamen 361004， China； 2.Department of Neurosurgery， Zhongshan Hospital Affiliated to Xiamen University， Fujian Province， Xiamen 361004， China 　　[Abstract] Objective To investigate the property of TTA1-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. TTA1-Tat-PEG-GS NPs bound to siRNA via electrostatic interaction. The composites of experiment included control group， TTA1-Tat-PEG-GS NPs/siRNA-Scr group， TTA1-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. Real-time 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 TTA1-Tat-P