Silencing of the scavenger receptor (Class B - Type 1) gene using siRNA-loaded chitosan nanaoparticles in a HepG2 cell model
Silencing of the scavenger receptor (Class B - Type 1) gene using siRNA-loaded chitosan nanaoparticles in a HepG2 cell model
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DOI：
10.1016/j.colsurfb.2014.10.045
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发表时间：
2014-11-01
影响因子：
5.8
通讯作者：
Abou-Aisha, Khaled
中科院分区：
文献类型：
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作者：
Farid, Mariane M.;Hathout, Rania M.;Abou-Aisha, Khaled
小干扰RNA （siRNA）介导的基因沉默在过去几十年里得到了越来越多的关注。然而，部分负电荷和易被核酸酶降解阻碍了其裸形式的使用。在这项研究中，我们研究了壳聚糖纳米颗粒作为siRNA的非病毒递送载体的使用。作为模型靶点，我们选择了清道夫受体（SR-B1），因为它被认为参与丙型肝炎病毒（HCV）的内化。以三聚磷酸钠（TPP）为交联剂，采用简单离子凝胶法制备了低分子量壳聚糖纳米颗粒；壳聚糖和TPP的浓度为0.1%，壳聚糖与TPP的质量比分别为3:1、5:1和9:1。采用fitc标记的壳聚糖纳米颗粒测定纳米颗粒的摄取效率，并采用Western blot检测HepG2细胞株中清除受体B类1 （SR-B1）的沉默。制备的纳米颗粒呈球形，具有最佳的粒径和分布。研究发现，HepG2细胞对fitc标记纳米颗粒的摄取具有浓度和时间依赖性。此外，Western Blot分析显示，SR-B1 siRNA能够在HepG2细胞中沉默清道夫受体长达96小时。(C) 2014 Elsevier B.V.版权所有
Gene silencing mediated by small interfering RNA (siRNA) has gained increasing interest through the past few decades. However, the partial negative charge and the susceptibility to degradation by nucleases have hampered its use in a naked form. In this study, we investigated the use of chitosan nanoparticles as non-viral delivery carriers of siRNA. As a model target, we selected the scavenger receptor (SR-B1), due to its proposed involvement in hepatitis C virus (HCV) internalization. Low molecular weight (LMW) chitosan nanoparticles were prepared by simple ionic gelation using sodium tripolyphosphate (TPP) as a cross-linking agent; a fixed chitosan and TPP concentration of 0.1% was used, and a chitosan to TPP weight ratios of 3:1, 5:1, and 9:1 were investigated. Nanoparticle uptake efficiency was measured using FITC-labeled chitosan nanoparticles and silencing of scavenger receptor class B type 1 (SR-B1) in HepG2 cell line was tested using Western blot analysis. Nanoparticles produced were spherical in shape with an optimum particle size and distribution. The uptake of FITC-labeled nanoparticles by HepG2 cells was found to be both concentration and time dependent. Furthermore, Western Blot analysis showed that SR-B1 siRNA was able to silence the scavenger receptor for up to 96 h of incubation with HepG2 cells. (C) 2014 Elsevier B.V. All rights reserved.