Efficient siRNA delivery and tumor accumulation mediated by ionically cross-linked folic acid-poly(ethylene glycol)-chitosan oligosaccharide lactate nanoparticles: For the potential targeted ovarian cancer gene therapy
Efficient siRNA delivery and tumor accumulation mediated by ionically cross-linked folic acid-poly(ethylene glycol)-chitosan oligosaccharide lactate nanoparticles: For the potential targeted ovarian cancer gene therapy
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DOI：
10.1016/j.ejps.2013.10.011
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发表时间：
2014-02-14
影响因子：
4.6
通讯作者：
Honke, Koichi
中科院分区：
文献类型：
--
作者：
Li, Tony Shing Chau;Yawata, Toshio;Honke, Koichi
为了有效地进行卵巢癌基因治疗，全身施用肿瘤靶向siRNA/叶酸聚（乙二醇）-壳寡糖乳酸（FA-PEG-COL）纳米颗粒对于递送到癌症部位是至关重要的。通过离子凝胶法制备siRNA/FA-PEG-COL纳米粒，用于有效转染表达FA受体的卵巢癌细胞并在体内蓄积。采用MALDI-TOF-MS、FT-IR和1H-1 NMR等手段对FAPEG-COL偶联物的结构进行了表征。siRNA/FA-PEG-COL纳米颗粒的平均尺寸约为200 nm，表面电荷为+8.4 mV，而siRNA/COL纳米颗粒为+30.5 mV。FA-PEG-COL纳米颗粒在聚集程度和溶血活性方面表现出与红细胞的上级相容性，并且与COL纳米颗粒相比，对细胞活力的影响也较低。如流式细胞术所示，FA接枝通过受体介导的内吞作用显著促进纳米颗粒的摄取。HIF-1 α的体外转染和基因敲低效率上级COL纳米颗粒（分别为76- 62%），并且与Lipofectamine 2000（79%）相当，如通过RT-qPCR和Western印迹所证明的。在分子水平上的基因敲除转化为体外增殖的有效抑制。使用体内成像在携带OVK18#2肿瘤异种移植物的BALB/c小鼠中研究FA-PEG-COL纳米颗粒的累积效率。主动靶向FA-PEG-COL纳米颗粒显示出比被动靶向COL纳米颗粒显著更大的蓄积。基于所获得的结果，siRNA/FA-PEG-COL纳米颗粒显示出通过基因治疗有效治疗卵巢癌的潜力。(C)2013爱思唯尔有限公司版权所有。
For effective ovarian cancer gene therapy, systemic administrated tumor-targeting siRNA/folic acidpoly(ethylene glycol)-chitosan oligosaccharide lactate (FA-PEG-COL) nanoparticles is vital for delivery to cancer site(s). siRNA/FA-PEG-COL nanoparticles were prepared by ionic gelation for effective FA receptor-expressing ovarian cancer cells transfection and in vivo accumulation. The chemical structure of FAPEG-COL conjugate was characterized by MALDI-TOF-MS, FT-IR and H-1 NMR. The average size of siRNA/FA-PEG-COL nanoparticles was approximately 200 nm, and the surface charge was +8.4 mV compared to +30.5 mV with siRNA/COL nanoparticles. FA-PEG-COL nanoparticles demonstrated superior compatibility with erythrocytes in terms of degree of aggregation and haemolytic activity and also effects on cell viability was lower when compared with COL nanoparticles. FA grafting significantly facilitated the uptake of nanoparticles via receptor mediated endocytosis as demonstrated by flow cytometry. The in vitro transfection and gene knockdown efficiency of HIF-1 alpha were superior to COL nanoparticles (76-62%, respectively) and was comparable to Lipofectamine 2000 (79%).as demonstrated by RT-qPCR and Western blot. Gene knockdown at the molecular level translated into effective inhibition of proliferation in vitro. Accumulation efficiency, of FA-PEG-COL nanoparticles was investigated in BALB/c mice bearing OVK18 #2 tumor xenograft using in vivo imaging. The active targeting FA-PEG-COL nanoparticles showed significantly greater accumulation than the passive targeting COL nanoparticles. Based on the results obtained, siRNA/FA-PEG-COL nanoparticles show much potential for effective ovarian cancer treatment via gene therapy. (C) 2013 Elsevier B.V. All rights reserved.