Hadianamrei Roja, Tomeh Mhd Anas, Wang Jiqian, Brown Stephen, Zhao Xiubo
Department of Chemical and Biological Engineering, University of Sheffield, S1 3JD, UK; School of Pharmacy and Biomedical Science, University of Portsmouth, PO1 2UP, UK.
Department of Chemical and Biological Engineering, University of Sheffield, S1 3JD, UK.
Biochem Biophys Res Commun. 2023 Apr 16;652:35-45. doi: 10.1016/j.bbrc.2023.02.026. Epub 2023 Feb 13.
Surfactant like peptides (SLPs) are a class of amphiphilic peptides widely used for drug delivery and tissue engineering. However, there are very few reports on their application for gene delivery. The current study was aimed at development of two new SLPs, named (IA)K and (IG)K, for selective delivery of antisense oligodeoxynucleotides (ODNs) and small interfering RNA (siRNA) to cancer cells. The peptides were synthesized by Fmoc solid phase synthesis. Their complexation with nucleic acids was studied by gel electrophoresis and DLS. The transfection efficiency of the peptides was assessed in HCT 116 colorectal cancer cells and human dermal fibroblasts (HDFs) using high content microscopy. The cytotoxicity of the peptides was assessed by standard MTT test. The interaction of the peptides with model membranes was studied using CD spectroscopy. Both SLPs delivered siRNA and ODNs to HCT 116 colorectal cancer cells with high transfection efficiency which was comparable to the commercial lipid-based transfection reagents, but with higher selectivity for HCT 116 compared to HDFs. Moreover, both peptides exhibited very low cytotoxicity even at high concentrations and long exposure time. The current study provides more insights into the structural features of SLPs required for nucleic acid complexation and delivery and can therefore serve as a guide for the rational design of new SLPs for selective gene delivery to cancer cells to minimize the adverse effects in healthy tissues.
类表面活性剂肽（SLPs）是一类两亲性肽，广泛应用于药物递送和组织工程。然而，关于其在基因递送方面的应用报道却非常少。当前的研究旨在开发两种新的SLPs，即(IA)K和(IG)K，用于将反义寡脱氧核苷酸（ODNs）和小干扰RNA（siRNA）选择性递送至癌细胞。这些肽通过Fmoc固相合成法合成。通过凝胶电泳和动态光散射研究了它们与核酸的络合情况。使用高内涵显微镜在HCT 116结肠癌细胞和人皮肤成纤维细胞（HDFs）中评估了这些肽的转染效率。通过标准MTT试验评估了这些肽的细胞毒性。使用圆二色光谱研究了这些肽与模型膜的相互作用。两种SLPs都能将siRNA和ODNs高效递送至HCT 116结肠癌细胞，其转染效率与市售脂质体转染试剂相当，但与HDFs相比，对HCT 116具有更高的选择性。此外，即使在高浓度和长时间暴露的情况下，这两种肽的细胞毒性都非常低。当前的研究为核酸络合和递送所需的SLPs结构特征提供了更多见解，因此可为合理设计新的SLPs以选择性地将基因递送至癌细胞以最小化对健康组织的不良影响提供指导。
