阳离子磷酸胆碱聚合物负载siRNA进行细胞转染的可行性  

Feasibility of siRNA delivery with cationic phosphorylcholine polymer in cells

作　　者：孙莉莉[1] 张卓琦[1] 曹希传[1] 杨帆[1] 李秀华[1] 潘修成[1] 

机构地区：[1]徐州医学院附属医院感染病科,221004

出　　处：《江苏医药》2010年第13期1570-1573,共4页Jiangsu Medical Journal

基　　金：国家自然科学基金(30800219);江苏省自然科学基金(BK2008132)

摘　　要：目的探讨新型阳离子磷酸胆碱聚合物(MPC30DEA70)负载短片断干扰RNA(siRNA)进行细胞转染的可行性以及生物学效应。方法 MTT法分析MPC30DEA70对HepG2.2.15细胞的毒性;设计特异性siRNA,以MPC30DEA70负载红色荧光标记的siRNA转染HepG2.2.15细胞,在荧光显微镜下观察该载体携带siRNA能否进入细胞中及在该细胞中的转染效率。在此基础之上,观察对荧光素酶基因表达的抑制效应。结果在MPC30DEA70的浓度为0.5、1、3、5μl时,未发现对HepG2.2.15细胞产生明显的毒性,细胞存活率均在80%以上,并且能够携带siRNA进入细胞内,在48h的转染效率达(52.33±1.97)%,转染后能够有效地抑制荧光素酶质粒在HepG2.2.15细胞内的表达。结论 MPC30DEA70携带的siRNA在HepG2.2.15细胞中有着较高的转染效率,并发挥一定的生物效应,有望成为有效的siRNA载体。Objective To observe the potential of a cationic phosphorylcholine polymer（MPC30DEA70）,a novel cationic phosphorylcholine polymer as a small interference RNA（siRNA） carrier and post-transcriptional gene silencing.Methods Cytotoxicity of MPC30DEA70 was evaluated by MTT colorimetric assay in HepG2.2.15 cells.After establishing the special siRNA,MPC30DEA70 carrying siRNA without and with red fluorescence transfected HepG2.2.15 cells,the efficiency of transfection was observed by fluorescence microscope.The effect of suppression was measured with luciferase assay system.Results Cytotoxicity of MPC30DEA70 was not apparent in the concentrations of 0.5,1,3,5μl,the efficiency of cell survival was greater than 80%,and MPC30DEA70 complexes could be transfected into HepG2.2.15 cells lines.These provided the level of transfection（52.33±1.97）% after 48 h and suppressed the expression of luciferase plasmid effectively in the cells.Conclusion MPC30DEA70 provides a higher level of transfection in HepG2.2.15 cells and produces some biological effect,which could be a carrier for siRNA delivery in the future.

关 键 词：阳离子磷酸胆碱聚合物 短片断干扰RNA 细胞转染 

分 类 号：R392[医药卫生—免疫学]