新型骨靶向纳米复合物抗前列腺癌骨转移细胞增殖作用研究

The study of new targeted-delivery of micro RNAs to bone-metastatic prostate tumors

目的 通过构建可携带具有抗前列腺癌增殖作用的基因miR-15-a和miR-16-1的适体-去端胶原（APT-ATE）,探讨载体系统的转染效率及其对前列腺癌细胞的抑制作用.方法 运用流式细胞仪,得出APT-ATE基因载体结构;利用粒径电位仪对APT-ATE/miRNA（适体-去端胶原/微小RNA）纳米复合物进行表征;利用基因转染考察APT-ATE/miRNA复合物在前列腺癌细胞（PC3和LNCaP）上的表达效果;CCK-8增殖抑制实验,考察APT-ATE/miRNA对前列腺癌细胞的抑制效果.结果 利用结构鉴定证明APT-ATE合成成功.定性定量转染效率实验证明APT修饰后的ATE对LNCaP细胞转染效率显著增加,证明APT的靶向作用.CCK-8细胞增殖实验证明,APT-ATE/miRNA可抑制前列腺癌细胞.裸鼠体内靶向性实验证明,APT-ATE/miRNA具有较好的骨靶向作用.结论 APT-ATE/miRNA有望成为今后靶向治疗骨转移性前列腺癌的基因药物.

Objective To construct a new gene delivery system based on atelocollagen （ATE）,and explore that modified aptamer （APT）,and APT-ATE/miRNA （miRNA-15a and miRNA-16-1） were successfully synthesized to treat bone-metastatic prostatic cancers.Methods Flow cytometry （FCM） analysis was used to characterize APT-ATE complex.The diameter and zeta potential of complexes were measured by Zetasizer Nano-ZS9.The prostatic cancer （PCa） distribution experiments were used to explore its biological characteristics and targeting ability of PCa cells （PC3 and LNCaP）.The inhibition of APT-ATE complex on LNCaP cell was determined with the cholecystokinin （CCK）-8 assay.Results FCM results demonstrated the successful synthesis of ATE-APT complex.The cellular uptake of vectors was concentration-dependent.The gene expression in vitro indicated that the modification of APT could increase the efficiency of gene expression and PCa targeting ability of ATE vectors to LNCaP [prostate specific membrane antigen （PSMA） over-expressing prostate cancer cells].The result of biodistribution showed that the bone uptake of APT-ATE was higher than ATE-APT.Conclusions APT-ATE/miRNA might be useful for preclinical and clinical studies on the treatment of bone-metastatic PCa.