Cellular uptake, distribution and cytotoxicity of the hydrophobic cell penetrating peptide sequence PFVYLI linked to the proapoptotic domain peptide PAD
Cellular uptake, distribution and cytotoxicity of the hydrophobic cell penetrating peptide sequence PFVYLI linked to the proapoptotic domain peptide PAD
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DOI：
10.1016/j.jconrel.2009.04.028
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发表时间：
2009-12-16
影响因子：
10.8
通讯作者：
Jones, Arwyn T.
中科院分区：
文献类型：
--
作者：
Watkins, Catherine L.;Brennan, Paul;Jones, Arwyn T.
细胞穿透肽（CPP）突破生物膜的能力为它们作为载体用于递送小分子药物和大分子治疗剂提供了希望。使用三种不同的细胞系统，包括原代人细胞，我们研究了充分表征的八精氨酸和最近发现的疏水性PFVYLI肽单独或与促凋亡结构域肽PAD（klaklak）缀合的摄取、亚细胞定位和对细胞活力的影响（2）。八精氨酸和PFVYLI在37 ℃下被有效地内吞到细胞中，但是在较高肽浓度下或在冰上进行摄取实验时直接穿过质膜的能力被限制在阳离子变体中。八精氨酸-和PFVYLI-PAD缀合物具有细胞毒性，其中KG 1a白血病细胞比HeLa细胞更敏感，并且八精氨酸-PAD是两种细胞系中最有效的缀合物。CPP-PAD缀合物对细胞形态和渗透性的影响是快速的，表明细胞毒性部分在质膜处介导，而不是完全通过诱导线粒体处的细胞凋亡。原代人白血病细胞与KG 1a细胞比HeLa细胞更相似，这表明白血病细胞对这些肽的相对敏感性可以在体内利用。(C)2009 Elsevier B. V.保留所有权利。
The capacity of cell penetrating peptides (CPPs) to breach biological membranes offers hope for their utilisation as vectors for the delivery of small molecule drugs and macromolecular therapeutics. Using three different cell systems, including primary human cells, we have studied the uptake, subcellular localisation and effect on cell viability of the well characterised octaarginine and the more recently discovered hydrophobic PFVYLI peptide, either alone, or conjugated to the proapoptotic domain peptide PAD (klaklak)(2). Octaarginine and PFVYLI were efficiently endocytosed into cells at 37 degrees C but an ability to translocate directly across the plasma membrane at higher peptide concentrations or when uptake experiments were performed on ice was confined to the cationic variant. Octaarginine- and PFVYLI-PAD conjugates were cytotoxic, with KG1a leukaemia cells being more sensitive than HeLa cells and octaarginine-PAD being the most potent conjugate in both cell lines. The effects of the CPP-PAD conjugates on cell morphology and permeability was rapid suggesting that cytotoxicity is partially mediated at the plasma membrane rather than exclusively through induction of apoptosis at the mitochondria. Primary human leukaemia cells were more similar to KG1a cells than HeLa cells, suggesting the relative sensitivity of leukaemia cells to these peptides could be exploited in vivo. (C) 2009 Elsevier B.V. All rights reserved.