Self-Assembled DNA Nanostructure as a Carrier for Targeted siRNA Delivery in Glioma Cells.
Self-Assembled DNA Nanostructure as a Carrier for Targeted siRNA Delivery in Glioma Cells.
复制标题
自组装 DNA 纳米结构作为胶质瘤细胞中靶向 siRNA 递送的载体
DOI：
10.2147/ijn.s295598
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发表时间：
2021
影响因子：
8
通讯作者：
Cheng Y
中科院分区：
文献类型：
--
作者：
Zhou Y;Yang Q;Wang F;Zhou Z;Xu J;Cheng S;Cheng Y
RNA干扰是一种很有前途的胶质瘤治疗方法。然而，由于RNA的不稳定性和缺乏肿瘤靶向性，限制了RNA干扰技术在胶质瘤治疗中的应用。在这里，我们报告了一种新的DNA四面体，它可以有效地提供小干扰RNA到胶质瘤细胞和诱导凋亡。方法将抑制Survivin表达的siRNA负载于DNA四面体（TDN）中。为了增强纳米粒子的主动靶向能力，我们在纳米粒子的一侧修饰了核酸适体as 1411（As-TDN-R），使其能够选择性地识别肿瘤细胞膜上的核仁素。通过琼脂糖凝胶电泳、动态光散射和透射电子显微镜对修饰后的纳米粒子进行了表征。琼脂糖凝胶电泳评价血清稳定性。免疫印迹和免疫荧光检测核仁素，并通过流式细胞术检测靶向细胞摄取。采用TUNEL法、流式细胞术和Western Blot法检测U87细胞凋亡情况。通过qPCR、Western Blot和免疫荧光检查存活素的基因沉默。结果As-TDN-R对siRNA具有较好的稳定性，表明TDN是一种良好的siRNA保护剂。与单独TDN相比，U87细胞对As-TDN-R的细胞间摄取增加，证明在胶质瘤细胞系中核仁素过表达。TUNEL法、流式细胞术和Western Blot显示As-TDN-R组细胞凋亡增加。Survivin蛋白和mRNA表达水平下调表明As-TDN-R有效地沉默了目的基因。结论该纳米粒可作为一种良好的siRNA靶向载体应用于胶质瘤的治疗。对DNA纳米结构的深入研究将极大地促进基于DNA的药物系统在胶质瘤中的应用。
Introduction RNA interference is a promising therapy in glioma treatment. However, the application of RNA interference has been limited in glioma therapy by RNA instability and the lack of tumor targeting. Here, we report a novel DNA tetrahedron, which can effectively deliver small interfering RNA to glioma cells and induce apoptosis. Methods siRNA, a small interfering RNA that can suppress the expression of survivin in glioma, was loaded into the DNA tetrahedron (TDN). To enhance the ability of active targeting of this nanoparticle, we modified one side of the DNA nanostructure with aptamer as1411 (As-TDN-R), which can selectively recognize the nucleolin in the cytomembrane of tumor cells. The modified nanoparticles were characterized by agarose gel electrophoresis, dynamic light scattering, and transmission electron microscopy. The serum stability was evaluated by agarose gel electrophoresis. Nucleolin was detected by Western blot and immunofluorescence, and targeted cellular uptake was examined by flow cytometry. The TUNEL assay, flow cytometry, and Western Blot were used to detect apoptosis in U87 cells. The gene silencing of survivin was examined by qPCR, Western Blot, and immunofluorescence. Results As-TDN-R alone showed better stability towards siRNA, indicating that TDN was a good siRNA protector. Compared with TDN alone, there was increased intercellular uptake of As-TDN-R by U87 cells, evidenced by overexpressed nucleolin in glioma cell lines. TUNEL assay, flow cytometry, and Western Blot revealed increased apoptosis in the As-TDN-R group. The downregulation of survivin protein and mRNA expression levels indicated that As-TDN-R effectively silenced the target gene. Conclusion The novel nanoparticle can serve as a good carrier for targeting siRNA delivery in glioma. Further exploration of the DNA nanostructure can greatly promote the application of DNA-based drug systems in glioma.