Cell line-dependent internalization pathways determine DNA transfection efficiency of decaarginine-PEG-lipid
Cell line-dependent internalization pathways determine DNA transfection efficiency of decaarginine-PEG-lipid
复制标题
DOI：
10.1016/j.ijpharm.2010.11.017
复制
发表时间：
2011-02-14
影响因子：
5.8
通讯作者：
Maitani, Yoshie
中科院分区：
文献类型：
--
作者：
Izumisawa, Tomohiro;Hattori, Yoshiyuki;Maitani, Yoshie
此前，我们曾报道十精氨酸缀合的 PEG-脂质 (R10B) 通过巨胞饮作用有效地将质粒 DNA (pDNA) 递送至人宫颈癌 HeLa 细胞中；然而，R10B 的细胞摄取机制尚未在其他细胞系中进行评估。在本研究中，我们研究了R10B/pDNA复合物（R10B-lipoplex）在人前列腺肿瘤PC-3和人鼻咽肿瘤KB细胞中的内化机制，并与HeLa细胞中的内化机制进行了比较。尽管在所有细胞系中，R10B-lipoplex 都必须与细胞表面的硫酸乙酰肝素 (HS) 结合，但 R10B-lipoplex 主要通过 PC-3 和 KB 细胞中的网格蛋白介导的内吞作用以及 HeLa 细胞中的巨胞饮作用来内化。在 HeLa 细胞中，用 R10B-lipoplex 处理可诱导用于巨胞饮作用的片状伪足的形成，但在 KB 和 PC-3 细胞中则不然。此外，在 HeLa 细胞中观察到 R10B-lipoplex 的转染效率最高。这些发现表明，R10B-lipoplex 与细胞上的 HS 结合后，诱导 HeLa 细胞中板状伪足的形成，然后通过巨胞饮作用内化，由于逃避了溶酶体中的降解，因此可以诱导高基因表达。细胞生理学可能是细胞内化和细胞穿透肽有效转染的关键因素。 (C) 2010 Elsevier B.V. 保留所有权利。
Previously, we have reported that decaarginine-conjugated PEG-lipids (R10B) efficiently delivered plasmid DNA (pDNA) into human cervical carcinoma HeLa cells via macropinocytosis; however, the mechanism of cellular uptake by R10B was not evaluated in other cell lines. In this study, we investigated the internalization mechanism by R10B/pDNA complex (R10B-lipoplex) in human prostate tumor PC-3 and human nasopharyngeal tumor KB cells, and compared with that in HeLa cells. Although it was necessary for R10B-lipoplex to associate with heparan sulfate (HS) on the cell surface in all cell lines, the R10B-lipoplex was internalized primarily through clathrin-mediated endocytosis in PC-3 and KB cells, and macropinosytosis in HeLa cells. In HeLa cells, treatment with the R10B-lipoplex induced the formation of lamellipodia for macropinocytosis, but did not in KB and PC-3 cells. Furthermore, the highest transfection efficiency by R10B-lipoplex was observed in HeLa cells. These findings indicated that the R10B-lipoplex induced the formation of lamellipodia in HeLa cells after binding to HS on the cells and was then internalized by macropinocytosis, which could induce high gene expression because of escaping degradation in lysosomes. Cell physiology might be a critical factor in cellular internalization and efficient transfection by cell penetration peptide. (C) 2010 Elsevier B.V. All rights reserved.