Facile Construction of Chloroquine Containing PLGA-Based pDNA Delivery System for Efficient Tumor and Pancreatitis Targeting in Vitro and in Vivo.
Facile Construction of Chloroquine Containing PLGA-Based pDNA Delivery System for Efficient Tumor and Pancreatitis Targeting in Vitro and in Vivo.
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DOI：
10.1021/acs.molpharmaceut.5b00155
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发表时间：
2015-05
影响因子：
4.9
通讯作者：
Chengli Yang;Tingting Hu;Hua Cao;Lijing Zhang;Pengxiang Zhou;Gu He;Xiangrong Song;A. Tong
中科院分区：
文献类型：
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作者：
Chengli Yang;Tingting Hu;Hua Cao;Lijing Zhang;Pengxiang Zhou;Gu He;Xiangrong Song;A. Tong
在传统的磷酸钙共沉淀法中，采用二磷酸氯喹（CQ）代替磷酸盐进行pDNA转染。由于CQ在新型Ca-CQ-pDNA复合物中的多重作用，包括pDNA压缩和协助溶酶体逃逸，pDNA的转染效率相对于传统方法显著增加。CQ没有像游离CQ那样插入DNA双螺旋中，这可能归因于pDNA与高浓度氯化钙的预先混合。为了构建体内基因传递载体，设计并制备了Ca-CQ-pDNA-PLGA-NPs。以包封率、粒径和pDNA完整性为筛选条件，得到最佳处方，并以优化后的磷酸钙共沉淀法制备的CaPi-pDNA-PLGA-NPs为对照，系统研究CQ在新型载体中的作用。利用TEM、DSC、XRD等手段对载体的物理性质进行了综合研究。采用MTT法、溶血试验和组织切片法评价了载体在体内外的安全性。与CaPi-pDNA-PLGA-NPs相比，Ca-CQ-pDNA-PLGA-NPs显著提高了人胚肾HEK 293细胞的基因转染效率，并在144 h内表现出增加的基因转染率。与来自纳米颗粒的pDNA相比，CQ的相对快速释放响应于增加的转染。在CT 26小鼠模型中，Did标记的-Ca-CQ-pDNA-PLGA-NPs表现出良好的肿瘤靶向效率和持续循环时间。将表达mSurvivin-T34 A蛋白的质粒pVITRO 2负载于Ca-CQ-pDNA-PLGA-NP中，抑瘤率为70%，这部分归因于CQ。Ca-CQ-pDNA-PLGA-NPs在C57急性胰腺炎模型中显示出较高的靶向性。总之，Ca-CQ-pDNA-PLGA-NP是肿瘤和胰腺炎靶向基因递送的有希望的候选物。
Chloroquine diphosphate (CQ) was ingeniously used to take place of phosphate salt in traditional calcium phosphate coprecipitation method for pDNA transfection. With multiple roles of CQ in the novel Ca-CQ-pDNA complex including pDNA compaction and assistance in lysosome escape, the transfection efficiency of the pDNA was significantly increased relative to the traditional method. CQ did not intercalate into the DNA double helix as free CQ did, which was probably ascribed to the prior mixing of the pDNA with high concentration of calcium chloride. In order to construct efficacious vector for in vivo gene delivery, Ca-CQ-pDNA-PLGA-NPs was designed and prepared. With entrapment efficiency, particle size and pDNA integrity as screening conditions, the optimal prescription was obtained and CaPi-pDNA-PLGA-NPs made with classic calcium phosphate coprecipitation method after optimization was also prepared as control to systematically study the role of CQ in the novel vector. Physical characters of the vectors were comprehensively studied using TEM, DSC, and XRD. The safety of the vector both in vitro and in vivo was evaluated using MTT, hemolysis test, and histological sections. The Ca-CQ-pDNA-PLGA-NPs dramatically enhanced the gene tranfection efficiency in Human Embryonic kidney HEK293 cells compared with the CaPi-pDNA-PLGA-NPs and presented an increasing gene transfection for up 144 h. The relative fast release of the CQ compared with pDNA from the nanoparticles was responsive for the increased transfection. The Did-labeled-Ca-CQ-pDNA-PLGA-NPs exhibited excellent tumor targeting efficiency and sustained circulation time in CT26 mouse model. The Ca-CQ-pDNA-PLGA-NP loaded with the plasmid pVITRO2 expressing mSurvivin-T34A protein gave 70% tumor inhibition rate, which was partially ascribed to CQ. The Ca-CQ-pDNA-PLGA-NPs showed high targeting efficiency in C57 acute pancreatitis model. In all, the Ca-CQ-pDNA-PLGA-NP was a promising candidate for targeted gene delivery to both tumor and pancreatitis.