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评价基于两亲肽的蛋白载体在体外癌症研究中的应用。

Evaluation of the use of amphipathic peptide-based protein carrier for in vitro cancer research.

机构信息

Institute of Bioengineering and Nanotechnology, 31 Biopolis Way, The Nanos, #04-01, 138669 Singapore, Singapore.

出版信息

Biochem Biophys Res Commun. 2012 Mar 9;419(2):170-4. doi: 10.1016/j.bbrc.2012.01.134. Epub 2012 Feb 3.

Abstract

Intracellular delivery of proteins offers an alternative to genetic modification or siRNA transfection for functional studies of proteins in live cells, especially for studies in cancer cells for therapeutics development. However, lack of efficient and biocompatible delivery system has limited the use of protein for in vitro cancer research. In this study, we design and evaluate an amphipathic peptide RV24, composing of a hydrophobic domain for protein binding, a flexible linker, and a hydrophilic domain to facilitate cell penetration. When using β-galactosidase as a cargo protein for comparison with commercially available peptide- and lipid-based carriers, RV24 peptide provides up to 5-fold increase in quantity delivered into 3 different cancer cell lines. Green fluorescent protein could also be delivered rapidly within 4h and transduced up to 83% of tested cancer cell lines. Although having a cell penetrating domain, RV24 peptide did not compromise cell viability, morphology and granularity significantly. These findings suggest the feasibility of using biocompatible amphipathic peptide to efficiently deliver protein-based molecules intracellularly for in vitro cancer research.

摘要

细胞内蛋白质递送为在活细胞中对蛋白质进行功能研究提供了一种替代遗传修饰或 siRNA 转染的方法,特别是在癌症细胞中进行治疗开发的研究。然而,缺乏有效的和生物相容的递送系统限制了蛋白质在体外癌症研究中的应用。在这项研究中,我们设计并评估了一种两亲性肽 RV24,它由一个用于蛋白质结合的疏水结构域、一个柔性连接子和一个亲水结构域组成,以促进细胞穿透。当使用β-半乳糖苷酶作为货物蛋白与市售的肽和基于脂质的载体进行比较时,RV24 肽将多达 5 倍的数量递送到 3 种不同的癌细胞系中。绿色荧光蛋白也可以在 4 小时内迅速递送到多达 83%的测试癌细胞系中。尽管具有穿透细胞的结构域,但 RV24 肽并没有显著影响细胞活力、形态和颗粒度。这些发现表明,使用生物相容的两亲性肽将基于蛋白质的分子有效地递送到细胞内进行体外癌症研究是可行的。

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