| 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/miRNA-FAM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。 |
| 关键词: 微RNAs 前列腺肿瘤 基因疗法 基因递送系统 |
| DOI：10.3724/SP.J.1008.2015.00117 |
| 投稿时间：2014-07-10修订日期：2014-10-14 |
| 基金项目:国家自然科学基金 (81302212, 81372762, 81272819). |
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| MicroRNA delivery system inhibits proliferation of androgen-independent prostate cancer |
| YAO Chong1,WU Xin2,TAI Zong-guang1,ZHU Quan-gang3,WANG Xiao-yu1,ZHANG Li-juan1,GAO Shen1* |
(1. Department of Pharmacy, Changhai Hospital, Second Military Medical University, Shanghai 200433, China;
2. Department of Pharmaceutics, First People's Hospital of Shanghai, Shanghai Jiaotong University, Shanghai 200080, China;
3. Department of Pharmacy, Yueyang Hospital of Integrated Traditional Chinese and Western Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai 200437, China
*Corresponding author) |
| Abstract: |
| Objective To prepare a microRNA (miRNA) delivery system using the branched polyethyleneimine (BPEI), linear polyethylenimine (LPEI) and polyamidoamine dendrimers (PAMAM) loaded with miRNA-15a and 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-15a and miRNA-16-1 against PC3 cells, and PCR was used to analyze their inhibitory effect on expression of Bcl-2, Cylin D1 and Wnt3a gene 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-15a and miRNA-16-1 into PC3 cells and block Bcl-2, Cylin D1 and Wnt3a expression in PC3 cells. Conclusion BPEI, LPEI and PAMAM loaded with miRNA-15a and miRNA-16-1 can suppress proliferation of prostate cancer PC3 cells and block Bcl-2, Cylin D1 and Wnt3a expression. |
| Key words: microRNAs prostatic neoplasms gene therapy gene delivery system |
