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阳离子碳硅烷树枝状聚合物及其与 siRNA 复合物与 MCF-7 细胞的相互作用。

Interaction of Cationic Carbosilane Dendrimers and Their siRNA Complexes with MCF-7 Cells.

机构信息

Department of General Biophysics, Faculty of Biology and Environmental Protection, University of Lodz, 141/143 Pomorska St., 90-236 Lodz, Poland.

Molecular and Nanostructural Biophysics Laboratory, "Bionanopark" Ldt., 114/116 Dubois St., 93-465 Lodz, Poland.

出版信息

Int J Mol Sci. 2021 Jul 1;22(13):7097. doi: 10.3390/ijms22137097.

DOI:10.3390/ijms22137097
PMID:34281151
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8269323/
Abstract

The application of siRNA in gene therapy is mainly limited because of the problems with its transport into cells. Utilization of cationic dendrimers as siRNA carriers seems to be a promising solution in overcoming these issues, due to their positive charge and ability to penetrate cell membranes. The following two types of carbosilane dendrimers were examined: CBD-1 and CBD-2. Dendrimers were complexed with pro-apoptotic siRNA (Mcl-1 and Bcl-2) and the complexes were characterized by measuring their zeta potential, circular dichroism and fluorescence of ethidium bromide associated with dendrimers. CBD-2/siRNA complexes were also examined by agarose gel electrophoresis. Both dendrimers form complexes with siRNA. Moreover, the cellular uptake and influence on the cell viability of the dendrimers and dendriplexes were evaluated using microscopic methods and XTT assay on MCF-7 cells. Microscopy showed that both dendrimers can transport siRNA into cells; however, a cytotoxicity assay showed differences in the toxicity of these dendrimers.

摘要

siRNA 在基因治疗中的应用主要受到其进入细胞的问题的限制。利用阳离子树状聚合物作为 siRNA 的载体似乎是克服这些问题的一种很有前途的解决方案，这是由于其正电荷和穿透细胞膜的能力。本研究考察了两种类型的碳硅烷树状聚合物：CBD-1 和 CBD-2。树状聚合物与促凋亡的 siRNA（Mcl-1 和 Bcl-2）复合，并通过测量其 zeta 电位、圆二色性和与树状聚合物相关的溴化乙锭的荧光来表征复合物。还通过琼脂糖凝胶电泳检查了 CBD-2/siRNA 复合物。两种树状聚合物都可以与 siRNA 形成复合物。此外，还使用 MCF-7 细胞的显微镜方法和 XTT 测定法评估了树状聚合物和树状聚合物复合物对细胞摄取和细胞活力的影响。显微镜观察表明，两种树状聚合物都可以将 siRNA 转运到细胞内；然而，细胞毒性测定表明这些树状聚合物的毒性存在差异。

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