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脂质体转染试剂和树枝状聚合物用于将短RNA转染至人T47D和MCF-10A细胞系的比较评估

Comparative Evaluation of Lipofectamine and Dendrimer for Transfection of Short RNA Into Human T47D and MCF-10A Cell Lines.

作者信息

Jahanafrooz Zohreh, Bakhshandeh Behnaz, Shirzadi Erfan

机构信息

Department of Biology, Faculty of Sciences, University of Maragheh, Maragheh, Iran.

Department of Biotechnology, College of Science, University of Tehran, Tehran, Iran.

出版信息

Adv Pharm Bull. 2023 Mar;13(2):385-392. doi: 10.34172/apb.2023.022. Epub 2022 Apr 30.

DOI:10.34172/apb.2023.022
PMID:37342383
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10278214/
Abstract

Non-viral transfection approaches are extensively used in cancer therapy. The future of cancer therapy lies on targeted and efficient drug/gene delivery. The aim of this study was to determine the transfection yields of two commercially available transfection reagents (i.e. Lipofectamine 2000, as a cationic lipid and PAMAM G5, as a cationic dendrimer) in two breast cell lines: cancerous cells (T47D) and non-cancerous ones (MCF-10A). We investigated the efficiencies of Lipofectamine 2000 and PAMAM G5 for transfection/delivery of a labeled short RNA into T47D and MCF-10A. In addition to microscopic assessments, the cellular uptakes of the complexes (fluorescein tagged-scrambled RNA with Lipofectamine or PAMAM dendrimer) were quantified by flow cytometry. Furthermore, the safety of the mentioned reagents was assessed by measuring cell necrosis through the cellular PI uptake. Our results showed significantly better efficiencies of Lipofectamine compared to PAMAM dendrimer for short RNA transfection in both cell types. On the other hand, MCF-10A resisted more than T47D to the toxicity of higher concentrations of the transfection reagents. Altogether, our research demonstrated a route for comprehensive epigenetic modification of cancer cells and depicted an approach to efficient drug delivery, which eventually improves both short RNA-based biopharmaceutical industry and non-viral strategies in epigenetic therapy.

摘要

非病毒转染方法在癌症治疗中被广泛应用。癌症治疗的未来在于靶向且高效的药物/基因递送。本研究的目的是确定两种市售转染试剂（即作为阳离子脂质的Lipofectamine 2000和作为阳离子树枝状聚合物的PAMAM G5）在两种乳腺细胞系中的转染效率：癌细胞（T47D）和非癌细胞（MCF-10A）。我们研究了Lipofectamine 2000和PAMAM G5将标记的短RNA转染/递送至T47D和MCF-10A细胞的效率。除了显微镜评估外，还通过流式细胞术对复合物（荧光素标记的乱序RNA与Lipofectamine或PAMAM树枝状聚合物）的细胞摄取进行了定量。此外，通过测量细胞对PI的摄取来评估上述试剂的安全性，以检测细胞坏死情况。我们的结果表明，在两种细胞类型中，与PAMAM树枝状聚合物相比，Lipofectamine在短RNA转染方面的效率显著更高。另一方面，MCF-10A比T47D对更高浓度转染试剂的毒性具有更强的抵抗力。总之，我们的研究展示了一种对癌细胞进行全面表观遗传修饰的途径，并描绘了一种高效药物递送的方法，最终改善了基于短RNA的生物制药行业以及表观遗传治疗中的非病毒策略。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/066b/10278214/db6653902b17/apb-13-385-g007.jpg
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ACS Appl Bio Mater. 2021 Sep 20;4(9):7168-7175. doi: 10.1021/acsabm.1c00743. Epub 2021 Aug 23.
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PAMAM-cRGD mediating efficient siRNA delivery to spermatogonial stem cells.PAMAM-cRGD 介导有效的 siRNA 递送至精原干细胞。
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Systematic Screening of Commonly Used Commercial Transfection Reagents towards Efficient Transfection of Single-Stranded Oligonucleotides.
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Molecules. 2018 Oct 8;23(10):2564. doi: 10.3390/molecules23102564.
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Role of dendrimers in advanced drug delivery and biomedical applications: a review.树枝状大分子在先进药物递送和生物医学应用中的作用：综述
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An improved method for increasing the efficiency of gene transfection and transduction.一种提高基因转染和转导效率的改进方法。
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