Tat-conjugated synthetic macromolecules facilitate cytoplasmic drug delivery to human ovarian carcinoma cells
Tat-conjugated synthetic macromolecules facilitate cytoplasmic drug delivery to human ovarian carcinoma cells
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DOI：
10.1021/bc0255900
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发表时间：
2003-01-01
影响因子：
4.7
通讯作者：
Kopecek, J
中科院分区：
文献类型：
--
作者：
Nori, A;Jensen, KD;Kopecek, J
合成了N-（2-羟丙基）甲基丙烯酰胺（HPMA）共聚物-细胞穿透肽Tat偶联物，并利用共聚焦荧光显微镜和亚细胞分离技术研究了它们在A2780人卵巢癌细胞中的亚细胞分布。我们的数据表明，单个Tat分子通过非内吞和浓度独立的过程将这些偶联物运输到细胞质和细胞核。通过活细胞显微镜观察，摄取在3分钟内发生。相比之下，缺乏Tat肽的HPMA共聚物仅通过内吞作用被内化。此外，还首次证实了tat介导的聚合物结合抗癌药物阿霉素的细胞质递送。这些发现确立了克服细胞内大分子递送的主要细胞和亚细胞障碍的可行性，并为开发基于聚合物的细胞质递送治疗分子的系统带来了巨大的希望。
We have synthesized N-(2-hydroxypropyl)methacrylamide (HPMA) copolymer-cell penetrating peptide Tat conjugates and evaluated their subcellular distribution in A2780 human ovarian carcinoma cells by confocal fluorescence microscopy and subcellular fractionation. Our data indicate the transport of these conjugates by a single Tat molecule to both the cytoplasm and nucleus via a nonendocytotic and concentration independent process. The uptake was observed to occur within 3 min, as confirmed by live cell microscopy. In contrast, HPMA copolymers lacking the Tat peptide were internalized solely by endocytosis. For the first time, Tat-mediated cytoplasmic delivery of a polymer bound anticancer drug, doxorubicin, was also demonstrated. These findings establish the feasibility of overcoming major cellular and subcellular obstacles to intracellular macromolecular delivery and hold great promise for the development of polymer-based systems for the cytoplasmic delivery of therapeutic molecules.