A Highly Efficient siRNA Carrier of PBLG Modified Hyperbranched PEI
A Highly Efficient siRNA Carrier of PBLG Modified Hyperbranched PEI
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PBLG修饰超支化PEI的高效siRNA载体
DOI：
10.1002/mabi.200900249
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发表时间：
2009-12-08
影响因子：
4.6
通讯作者：
Chen, Xuesi
中科院分区：
文献类型：
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作者：
Chen, Jie;Tian, Huayu;Chen, Xuesi
为了寻找有效的非病毒基因载体来传递siRNA，设计并合成了一种通过将疏水聚（γ‐苄基-谷氨酸）片段（PBLG）接枝到超支化聚乙烯亚胺（PEI‐PBLG）上的共聚物。PEI‐PBLG可以有效地将siRNA传递到细胞中以沉默靶基因。与未修饰的PEI相比，PEI‐PBLG显著降低了细胞毒性。siRNA与PEI - PBLG复合物在稳定表达的荧光素酶CT26细胞中显示出显著的靶向荧光素酶基因敲除率（相对于未处理的细胞，在孵卵6小时后不改变培养基），而Lipofectamine™2000和未修饰的PEI分别只能达到57.92%和15.31%的敲除率。在稳定转染了荧光素酶基因的4T1细胞和短暂转染了荧光素酶基因的HeLa细胞中，siRNA与PEI - PBLG复合物也表明，它比未修饰的PEI和Lipofectamin™2000具有更大的基因沉默能力。采用流式细胞术定量检测载体/Alexa647标记siRNA的内化效率。PEI‐PBLG/Alexa647‐标记的siRNA的内化效率为52.67%，而PEI和Lipofectamine™2000分别为27.23%和37.91%。共聚焦激光扫描显微镜（CLSM）实验也表明PEI‐PBLG诱导的细胞摄取效率高于其他商业试剂。PEI‐PBLG被证明是一种有前景的siRNA载体，在癌症治疗中具有潜在的应用前景。
In search for effective non‐viral gene vectors for the delivery of siRNA, a copolymer was designed and synthesized by grafting hydrophobic poly(γ‐benzylL‐glutamate) segment (PBLG) to hyperbranched polyethylenimine (PEI‐PBLG). PEI‐PBLG could efficiently deliver siRNA to cells to silence the targeted gene. Markedly, PEI‐PBLG caused lower cytotoxicity in comparison to unmodified PEI. The siRNA complexed with PEI‐PBLG showed a remarkable knockdown (75.23% relative to untreated cells, without changing the medium after 6 h of incubation) of the targeted luciferase gene in stable expressing luciferase CT26 cells while the Lipofectamine™2000 and unmodified PEI could only achieve knockdown rates of 57.92% and 15.31%, respectively. The siRNA complexed with PEI‐PBLG also demonstrated that it had greater gene silencing ability than unmodified PEI and Lipofectamin™2000 in both 4T1 cells stably transfected with the luciferase gene and HeLa cells transiently transfected with the luciferase gene. The internalization efficiency of carrier/Alexa647‐labeled siRNA was quantified using flow cytometry. PEI‐PBLG/Alexa647‐labeled siRNA showed internalization efficiency of 52.67% while PEI and Lipofectamine™2000 demonstrated 27.23% and 37.91%, respectively. Confocal laser scanning microscopy (CLSM) assay also indicated that PEI‐PBLG induced higher cell uptake efficiency than other commercial reagents. PEI‐PBLG was shown to be a promising siRNA carrier with potential application in cancer therapy.