线性和支链聚乙烯亚胺纳米结构对a431细胞的细胞毒性影响。

BioImpacts : BI Pub Date : 2011-01-01 Epub Date: 2011-06-09 DOI:10.5681/bi.2011.004
Vala Kafil, Yadollah Omidi
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引用次数: 215

摘要

简介:聚乙烯亚胺(PEI)作为一种非病毒阳离子聚合物,在基因传递纳米系统中得到了广泛的应用。尽管许多研究强调了其通过诱导细胞凋亡/坏死对靶细胞的毒性作用,但仍有必要研究其对靶细胞的结构影响。方法:在本研究中,我们报道了25 kD线性和分支PEI(分别为LPEI和BPEI)在A431细胞中的细胞基因组影响,以解决诱导凋亡的可能机制。在40-50%的合流度下,A431细胞以推荐浓度暴露于PEI中4小时。24h后,采用MTT法、fitc标记膜联蛋白V流式细胞术和comet法检测细胞凋亡和DNA损伤。结果:流式细胞术评估显示,与线性PEI相比,BPEI可导致更大的内化,这也诱导了更大的细胞毒性。膜联蛋白V检测证实BPEI早期和晚期细胞凋亡,彗星检测检测到一定程度的DNA损伤。Western blot分析结果显示,akt激酶可能是受PEI影响的生物分子之一。结论:这些结果表明,尽管有akt激酶的诱导,BPEI仍能引起靶细胞的凋亡。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Cytotoxic impacts of linear and branched polyethylenimine nanostructures in a431 cells.

Introduction: Polyethylenimine (PEI), as a nonviral cationic polymer, has been widely used as gene delivery nanosystem. Although a number of investigations have highlighted its toxic impacts on target cells through induction of apoptosis/necrosis, still it is essential to look at its structural impacts on target cells.

Methods: In this current study, cytogenomic impacts of 25 kD linear and branched PEI (LPEI and BPEI, respectively) in A431 cells are reported to address possible mechanism for induction of apoptosis. At 40-50% confluency, A431 cells were exposed to PEI at a recommended concentration for 4 hr. After 24 hr, to detect apoptosis and DNA damage, the treated cells were subjected to MTT assay, FITC-labeled annexin V flow cytometry and comet assay.

Results: Flow cytometry assessments revealed that the BPEI can result in greater internalization than the linear PEI, which also induced greater cytotoxicity. Annexin V assay confirmed early and late apoptosis by BPEI, imposing somewhat DNA damage detected by comet assay. Western blot analysis resulted in induction of Akt-kinase which is possibly one of biomolecules affected by PEI.

Conclusion: These results highlight that, despite induction of Akt-kinase, the BPEI can elicit apoptosis in target cells.

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