MicroRNA基因递送系统抗激素非依赖型前列腺癌增殖作用研究  

MicroRNA delivery system inhibits proliferation of androgen-independent prostate cancer

作　　者：姚翀[1] 武鑫[2] 台宗光 朱全刚[3] 王晓宇[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,上海200433 [2]上海交通大学附属第一人民医院临床药学科,上海200080 [3]上海中医药大学附属岳阳医院药剂科,上海200437

出　　处：《第二军医大学学报》2015年第2期117-123,共7页Academic Journal of Second Military Medical University

基　　金：国家自然科学基金(81302212;81372762;81272819)~~

摘　　要：目的采用支化的聚乙烯亚胺(BPEI)、线型聚乙烯亚胺(LPEI)和树枝状聚合物聚酰胺-胺(PAMAM)与具有抑制激素非依赖型前列腺癌细胞PC3增殖的microRNA(miRNA)-15a和miRNA-16-1质粒结合,构建了miRNA基因递送系统。考察形成的3种纳米复合物的粒径、zeta电位、细胞内摄取,以及对PC3细胞的抑制作用。方法利用粒径分析仪测定形成的3种纳米复合物的粒径和电位,凝胶电泳实验测定3种材料对miRNA的结合能力;利用FAM标记的NC-miRNA考察PC3细胞对3种复合物的摄取情况;通过CCK8法测定3种材料与miRNA-15a和miRNA-16-1形成的复合物对PC3细胞的抑制作用;PCR法测定3种材料与miRNA-15a和miRNA-16-1结合后对PC3细胞中Bcl-2、Cylin D1和Wnt3a表达的阻滞作用。结果BPEI、LPEI和PAMAM与miRNA可结合形成稳定的纳米级复合物,在N/P=5时,PC3细胞对BPEI/miRNAFAM的摄取高于LPEI/miRNA-FAM和PAMAM/miRNA-FAM,差异具有统计学意义(P<0.05)。BPEI、LPEI和PAMAM都可以携带miRNA-15a和miRNA-16-1进入PC3细胞,并且阻滞PC3细胞中Bcl-2、Cylin D1和Wnt3a蛋白的表达。结论BPEI、LPEI和PAMAM包裹miRNA-15a和miRNA-16-1可对激素非依赖型前列腺癌细胞PC3细胞产生抑制作用,并可以有效阻滞PC3细胞中Bcl-2、Cylin D1和Wnt3a蛋白的表达。Objective To prepare a microRNA（miRNA）delivery system using the branched polyethyleneimine（BPEI）,linear polyethylenimine（LPEI）and polyamidoamine dendrimers（PAMAM）loaded with miRNA-15 aand miRNA-16-1,which can inhibit prostate cancer PC3 cell proliferation,and to examine the zeta potential,intracellular uptake,and the inhibition effect on PC3 cells of the three constructed nano-complexes.Methods Particle size analyzer was used to determine the size and potential of the three kinds of nano-complexes,and the miRNA affinity capability of them was determined by agarose gel electrophoresis retardation assay.The uptake efficiency of the nano-complexes by PC3 cells was examined by NC-miRNA labeled with FAM.CCK8 method was used to determine the inhibitory effect of the nano-complexes loaded with miRNA-15 a and miRNA-16-1against PC3 cells,and PCR was used to analyze their inhibitory effect on expression of Bcl-2,Cylin D1 and Wnt3agene in PC3 cells.Results BPEI,LPEI and PAMAM loaded with miRNA could form stable nano-complexes.When N/P=5,the intracellular uptake of BPEI/miRNA-FAM by PC3 cells was significantly higher than that of LPEI/miRNA-FAM and PAMAM/miRNA-FAM（P〈0.05）.BPEI,LPEI and PAMAM could all carry miRNA-15 aand miRNA-16-1into PC3 cells and block Bcl-2,Cylin D1 and Wnt3aexpression in PC3 cells.Conclusion BPEI,LPEI and PAMAM loaded with miRNA-15 aand miRNA-16-1can suppress proliferation of prostate cancer PC3 cells and block Bcl-2,Cylin D1 and Wnt3aexpression.

关 键 词：微RNAS 前列腺肿瘤 基因疗法 基因递送系统 

分 类 号：R737.25[医药卫生—肿瘤]