聚乙二醇修饰的聚精氨酸基因载体的构建及其体外评价  

Preparation of polyethylene glycol-modified polyarginine for gene delivery and in vitro evaluation

作　　者：台宗光 孙霖 朱全刚[1] 张玮[1,3] 王晓宇[1] 戴子渊[1] 张丽娟[1] 高申[1] 

机构地区：[1]第二军医大学长海医院药学部 [2]解放军92330部队医院 [3]解放军522医院药材科

出　　处：《第二军医大学学报》2014年第10期1088-1093,共6页Academic Journal of Second Military Medical University

基　　金：国家自然科学基金(81272819;81172514;81101658)~~

摘　　要：目的利用聚乙二醇(PEG)修饰非病毒基因载体聚精氨酸(PLR),考察PEG修饰对PLR的细胞毒性和PLR介导的RNA干扰效率的影响。方法利用1 HNMR鉴定合成的PLR-PEG的结构,确定PEG的修饰度,利用凝胶电泳表征载体对siRNA的包裹能力,在前列腺癌干细胞模型细胞(RC-92a/hTERT)上考察PLR-PEG的细胞毒性,考察PLR-PEG/siRNA复合物的细胞摄取及相关基因的干扰效率。结果通过结构鉴定确定PLR-PEG合成成功;细胞毒性实验表明PEG修饰可以降低PLR的毒性;PEG修饰会降低PLR/siRNA复合物的细胞摄取,高N/P时对细胞摄取的影响不大;PEG修饰也会降低PLR介导的RNA干扰效率,但PEG修饰度在一定范围内对干扰效率的影响比较小。结论 PEG修饰的PLR作为基因载体在前列腺癌干细胞的基因治疗中有一定的应用前景。Objective To modify polyargine（PLR）with polyethylene glycol（PEG）and to observe the effect of PEG modification on PLR cytotoxicity and efficiency of PLR-mediated RNA interference.Methods 1 HNMR was used to characterize PLR-PEG and gel electrophoresis was adopted to determine the siRNA-packing capacity of PLR-PEG.The cytotoxicity of PLRPEG,cellular uptake and RNA interference efficiency of PLR-PEG/siRNA complexes were investigated using prostate cancer stem-like cells（RC-92a/hTERT）.Results 1 HNMR results showed the successful synthesis of PLR-PEG.It was found that PEG modification decreased cytotoxicity of PLR and reduced cellular uptake of PLR/siRNA complexes,but the reduction of cellular uptake was limited when N/P was high.The modification also inhibited the efficiency of PLR-mediated RNA interference,but the influence of PEG modification was not notable within a certain range.Conclusion PEG-modified PLR may be a promising vector for gene therapy targeting prostate cancer stem cells.

关 键 词：聚精氨酸 聚乙二醇 基因载体 细胞毒性 基因干扰效率 前列腺癌干细胞 

分 类 号：R943.4[医药卫生—药剂学]