Polyethylenimine as a gene delivery tool in triple-negative breast cancer cell line and breast cancer stem cell model

Pub Date : 2022-07-01 DOI:10.18054/pb.v123i3-4.10472
M. Matovina, Steffi Lemmens, M. Kralj, K. Ester
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Abstract

Background and purpose: PEI is a cationic polymer able to neutralise negative DNA charges and to condense large genes which makes it suitable for gene delivery in human cells. Despite its low cost, simplicity of use and moderate toxicity, there is still room for broader usage and experimental adjustments, especially in cell lines that are difficult to transfect. In the presented research, we used PEI for the delivery of plasmid DNA into TNBC cell line SUM159 and breast CSC model HMLE-Twist.Material and methods: Cultured cells were transfected with GFP-expressing plasmid using both PEI and Lipofectamin. Transfection efficiency was determined by flow cytometry measurements of the intensity of the green fluorescence, while viability was determined by measuring intensity of the red fluorescence after propidium iodide staining. Results: In SUM159 and HMLE-Twist cells we obtain transfection efficiency between 30-40% using PEI, while cytotoxicity was generally low to moderate. PEI caused 10% of cell death in SUM159 and 20 % in HMLE-Twist. Transfection efficiency of PEI was comparable and even higher than the efficiency of the Lipofectamine in both SUM159 and HMLE-Twist. In mammary epithelia (control HMLE) we obtained only 20 % transfection efficiency using both carriers.Conclusions: We demonstrated for the first time that PEI represents a suitable nanocarrier for gene delivery into breast CSC model. We successfully transfected both breast CSC model HMLE-Twist and TNBC line SUM159. Since PEI is inexpensive and easy to use, we recommend it for further exploitations of these cell lines in TNBC research.
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聚乙烯亚胺在三阴性乳腺癌细胞系和乳腺癌干细胞模型中的基因传递工具
背景和目的:PEI是一种阳离子聚合物,能够中和负的DNA电荷,并凝聚大的基因,使其适合于人类细胞中的基因传递。尽管其成本低,使用简单,毒性适中,但仍有更广泛使用和实验调整的空间,特别是在难以转染的细胞系中。在本研究中,我们使用PEI将质粒DNA传递到TNBC细胞系SUM159和乳腺CSC模型hmel - twist中。材料和方法:用PEI和Lipofectamin分别转染表达gfp的质粒。转染效率通过流式细胞术测量绿色荧光强度来确定,而生存能力通过碘化丙啶染色后测量红色荧光强度来确定。结果:在SUM159和hmel - twist细胞中,我们使用PEI获得了30-40%的转染效率,而细胞毒性一般为低至中等。在SUM159中,PEI导致10%的细胞死亡,在HMLE-Twist中,PEI导致20%的细胞死亡。PEI的转染效率与Lipofectamine在SUM159和hmel - twist中的转染效率相当,甚至更高。在乳腺上皮(对照HMLE)中,两种载体的转染效率仅为20%。结论:我们首次证明PEI是一种合适的纳米载体,可以将基因传递到乳腺CSC模型中。我们成功转染了乳腺CSC模型HMLE-Twist和TNBC细胞系SUM159。由于PEI价格低廉且易于使用,我们建议将其用于TNBC研究中这些细胞系的进一步开发。
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