Intracellular distribution and internalization pathways of guanidinylated bioresponsive poly(amido amine)s in gene delivery
Intracellular distribution and internalization pathways of guanidinylated bioresponsive poly(amido amine)s in gene delivery
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基因传递中胍基化生物响应性聚酰胺胺的细胞内分布和内化途径
DOI：
10.1016/j.ajps.2018.02.008
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发表时间：
2018-03
影响因子：
10.2
通讯作者：
Ding Pingtian
中科院分区：
文献类型：
--
作者：
Zhang Jinmin;Wang Chunxi;Lu Mei;Xing Haonan;Yang Tianzhi;Cai Cuifang;Zhao Xiaoyun;Wei Minjie;Yu Jiankun;Ding Pingtian
通过盐酸胍（CAR）或氯己定（CHL）与N,N'-胱胺双丙烯酰胺（CBA）之间的迈克尔型加成反应合成了胍基化生物响应性聚酰胺胺聚合物CAR-CBA和CHL-CBA。先前的研究表明，这两种聚合物作为基因递送载体具有较高的转染效率。在这项研究中，我们研究了这两种聚合物在基因传递中的核仁定位能力和细胞内化途径。每种聚合物都会浓缩质粒 DNA (pDNA) 并形成纳米颗粒复合物，然后在 MCF-7 细胞中进行转染研究。细胞转染后，两种复合物均富集于核仁中，并且当转染于M期停滞的MCF-7细胞时，其转染效率显着提高。氯丙嗪会抑制CAR-CBA-pDNA的转染效率，接触CAR-CBA-pDNA后细胞内体会被破坏。对于CHL-CBA-pDNA，其转染效率不受研究中使用的三种内吞作用抑制剂的影响，并且CHL-CBA-pDNA对内体没有显示出影响。两种复合物转染细胞后，细胞乳酸脱氢酶释放和膜形态发生变化。结果表明，CAR-CBA和CHL-CBA聚合物均表现出良好的核仁定位能力。细胞阻滞于M期有利于转染。 CAR-CBA-pDNA 细胞内化参与网格蛋白介导的内吞途径，并逃离内体包埋，而 CHL-CBA-pDNA 的细胞摄取通过网格蛋白和小凹独立的机制发生。 (c) 2018 沉阳药科大学。由 Elsevier B.V. 出版。这是一篇遵循 CC BY-NC-ND 许可的开放获取文章。 (http://creativecommons.org/licenses/by-nc-nd/4.0/)
Guanidinylated bioresponsive poly(amido amine)s polymers, CAR-CBA and CHL-CBA, were synthesized by Michael-type addition reaction between guanidine hydrochloride (CAR) or chlorhexidine (CHL) and N,N'-cystaminebisacrylamide (CBA). Previous studies have shown that both polymers had high transfection efficiencies as gene delivery carriers. In this study, we investigated the nucleolus localization abilities and cellular internalization pathways of these two polymers in gene delivery. Each polymer condensed plasmid DNA (pDNA) and formed nanoparticle complexes, and then their transfection studies were performed in MCF-7 cells. Both complexes were found enriched in nucleolus after cellular transfection, and their transfection efficiencies were significantly improved when transfection was performed on MCF-7 cells arrested at M phase. The transfection efficiency of CAR-CBA-pDNA was inhibited by chlorpromazine, and cell endosomes were disrupted after being exposed to CAR-CBA-pDNA. In regards to CHL-CBA-pDNA, its transfection efficiency was not affected by three types of endocytosis inhibitors used in the study, and CHL-CBA-pDNA showed no effect on endosomes. Cellular lactate dehydrogenase release and membrane morphology were changed after cells were transfected by the two complexes. The results indicated that both CAR-CBA and CHL-CBA polymers demonstrated good nucleolus localization abilities. It was beneficial for transfection when cells were arrested at M phase. CAR-CBA-pDNA cellular internalization was involved with clathrin-mediated endocytosis pathway, and escaping from endosomal entrapment, while the cellular uptake of CHL-CBA-pDNA occurs via clathrin-and caveolae-independent mechanism. (c) 2018 Shenyang Pharmaceutical University. Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND license. (http://creativecommons.org/licenses/by-nc-nd/4.0/)
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影响因子：
3.6
作者：
Singer O;Verma IM
通讯作者：
Verma IM
内吞作用未插入：多种进入细胞的方法。
影响因子：
44.1
作者：
Kumari S;Mg S;Mayor S
通讯作者：
Mayor S
新型胍基化生物响应性聚酰胺胺，专为短发夹 RNA 递送而设计
影响因子：
8
作者：
Yu J;Zhang J;Xing H;Sun Y;Yang Z;Yang T;Cai C;Zhao X;Yang L;Ding P
通讯作者：
Ding P
影响因子：
16.1
作者：
R. Wattiaux;Nathanael Laurent;S. Coninck;M. Jadot
通讯作者：
R. Wattiaux;Nathanael Laurent;S. Coninck;M. Jadot
影响因子：
10.8
作者：
Nam, Hae Yun;Kwon, Seok Min;Jeong, Seo Young
通讯作者：
Jeong, Seo Young