M.M. Shemyakin and Yu.A. Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences, Moscow, Russia.
Chem Biol Drug Des. 2018 Jan;91(1):294-303. doi: 10.1111/cbdd.13081. Epub 2017 Sep 11.
Natural antimicrobial peptides (AMPs) are important components of the innate immune system with a wide spectrum of biological activity. In this study, we investigated the cytotoxic effect of three recombinant β-hairpin cationic AMPs: arenicin-1 from the polychaeta Arenicola marina, tachyplesin I from the horseshoe crab Tachypleus tridentatus, and gomesin from the spider Acanthoscurria gomesiana. All the three β-hairpin AMPs were overexpressed in Escherichia coli. Different cell lines were incubated with various concentrations of the investigated AMPs in order to evaluate their cytotoxic activity. Double staining with subsequent flow cytometric analysis was used to determine the predominant way of cell death mediated by each AMP. Hemolytic activity of the peptides was tested against fresh human red blood cells. Our results indicated that all the three AMPs exhibited significant cytotoxic effect against cancer cells that varied depending on the cell line type and, in most cases, on the presence of serum components. Flow cytometric analysis implicitly indicated that tachyplesin I mostly promoted late apoptosis/necrosis, while arenicin-1 and gomesin induced early apoptosis under the same conditions. Tachyplesin I proved to be the most promising therapeutic candidate as it displayed the highest specific cytotoxicity against cancer cell lines, independent of the serum presence.
天然抗菌肽 (AMPs) 是先天免疫系统的重要组成部分，具有广泛的生物活性。在本研究中，我们研究了三种重组 β-发夹阳离子 AMPs 的细胞毒性作用：来自多毛纲海洋生物沙蚕的海鞘素 I、来自鲎的鲎素 I 和来自蜘蛛的 gomesin。所有三种 β-发夹 AMP 均在大肠杆菌中过表达。用不同浓度的研究 AMP 孵育不同的细胞系，以评估它们的细胞毒性活性。通过双染色随后进行流式细胞术分析来确定每种 AMP 介导的细胞死亡的主要方式。用肽对新鲜的人红细胞进行溶血活性测试。我们的结果表明，所有三种 AMP 对癌细胞均表现出显著的细胞毒性作用，其作用因细胞系类型而异，并且在大多数情况下，还与血清成分的存在有关。流式细胞术分析隐含地表明，在相同条件下，鲎素 I 主要促进晚期细胞凋亡/坏死，而 arenicin-1 和 gomesin 诱导早期细胞凋亡。鲎素 I 表现出对癌细胞系的最高特异性细胞毒性，且不依赖于血清的存在，因此被证明是最有前途的治疗候选物。
