Multifunctional DNA-Gold Nanoparticles for Targeted Doxorubicin Delivery
Multifunctional DNA-Gold Nanoparticles for Targeted Doxorubicin Delivery
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DOI：
10.1021/bc500136r
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发表时间：
2014-07-25
影响因子：
4.7
通讯作者：
Dabrowiak, James C.
中科院分区：
文献类型：
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作者：
Alexander, Colleen M.;Hamner, Kristen L.;Dabrowiak, James C.
在这份报告中，我们描述了合成，表征和细胞毒性性质的DNA-帽的金纳米粒子连接叶酸（FA），热响应聚合物（P），和/或聚（乙二醇）（PEG）的低聚物，可用于提供抗癌药物阿霉素（DOX）在化疗。通过分离的叶酸N-羟基琥珀酰亚胺酯与氨基-DNA的反应合成用于构建递送载体的FA-DNA寡聚体，并使用高效液相色谱（HPLC）纯化缀合的DNA产物。这种方法最终允许控制附着于递送载体表面的FA的量。使用SK-N-SH神经母细胞瘤细胞和载药递送载体进行细胞毒性研究，使用各种暴露时间（1-48 h）和恢复时间（1-72 h），并在FA缺乏的培养基中获得不同量的附着FA的影响。具有50%的DNA链与附着的FA的DOX负载的递送载体比当所有链含有FA时更具有细胞毒性。由于FA刺激细胞生长，因此完全覆盖FA的溶媒的细胞毒性降低表明FA的刺激作用可超过药物对细胞群的细胞毒性作用。虽然将六乙二醇PEG（18）连接到递送载体的表面对细胞毒性没有影响，但100%FA加上温敏聚合物导致IC 50 = 0.48 +/- 0.01，暴露时间为24小时，恢复时间为1小时，其细胞毒性比游离DOX大一个数量级。使用荧光检测的共聚焦显微镜研究显示，暴露于装载DOX的载体的SK-N-SH神经母细胞瘤细胞在细胞内具有药物积累，并且在具有附着的FA和温敏聚合物的载体的情况下，药物似乎更浓缩。由于DOX的生物靶标是DNA，后一观察结果与具有FA和温敏聚合物的载体的高细胞毒性一致。这项研究强调了DNA加帽的金纳米颗粒作为癌症化疗中多柔比星递送载体的潜力。
In this report we describe the synthesis, characterization, and cytotoxic properties of DNA-capped gold nanoparticles having attached folic acid (FA), a therrnoresponsive polymer (p), and/or poly(ethylene glycol) (PEG) oligomers that could be used to deliver the anticancer drug doxorubicin (DOX) in chemotherapy. The FA-DNA oligomer used in the construction of the delivery vehicle was synthesized through the reaction of the isolated folic acid N-hydroxysuccinimide ester with the amino-DNA and the conjugated DNA product was purified using high performance liquid chromatography (HPLC). This approach ultimately allowed control of the amount of FA attached to the surface of the delivery vehicle. Cytotoxicity studies using SK-N-SH neuroblastoma cells with drug loaded delivery vehicles were carried out using a variety of exposure times (1-48 h) and recovery times (1-72 h), and in order to access the effects of varying amounts of attached FA, in culture media deficient in FA. DOX loaded delivery vehicles having 50% of the DNA strands with attached FA were more cytotcodc than when all of the strands contained FA. Since FA stimulates cell growth, the reduced cytotoxicity of vehicles fully covered with FA suggests that the stimulatory effects of FA can more than compensate for the cytotoxic effects of the drug on the cell population. While attachment of hexa-ethylene glycol PEG(18) to the surface of the delivery vehicle had no effect on c-ytotoxicity, 100% FA plus the thermoresponsive polymer resulted in IC50 = 0.48 +/- 0.01 for an exposure time of 24 h and a recovery time of 1 h, which is an order of magnitude more cytotoxic than free DOX. Confocal microscopic studies using fluorescence detection showed that SK-N-SH neuroblastoma cells exposed to DOX-loaded vehicles have drug accumulation inside the cell and, in the case of vehicles with attached FA and thermoresponsive polymer, the drug appears more concentrated. Since the biological target of DOX is DNA, the latter observation is consistent with the high cytotoxicity of vehicles having both FA and the thermoresponsive polymer. The study highlights the potential of DNA-capped gold nanoparticles as delivery vehicles for doxorubicin in cancer chemotherapy.