RNA interference is a powerful approach to understand gene function both for therapeutic and experimental purposes. Since the lack of knowledge in the gene silencing of various hepatic cell lines, this work was aimed to compare two transfection agents, the liposome-based Lipofectamine™ RNAiMAX and the HepG2-specific, polymer-based GenMute™, in two cellular models of human hepatoma, HepG2 and Huh7.5. In the first part, we assessed transfection efficiency of a fluorescent Cy3-labeled negative control siRNA by cell imaging analysis; we found that cells treated with GenMute present a higher uptake of the fluorescent negative control siRNA when compared to Lipofectamine RNAiMAX-transfected cells, both in HepG2 and in Huh7.5 cells. In the second part, we evaluated GAPDH silencing with the two transfection reagents by RT-PCR similar GAPDH mRNA expression after each transfection treatment. Finally, we measured cell viability by the MTT assay, observing that cells transfected with GenMute have higher viability with respect to Lipofectamine RNAiMAX-administered cells. These results suggest that GenMute reagent might be considered the most suitable transfection agent for hepatic gene silencing.

中文翻译：

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Lipofectamine RNAiMAX和GenMute转染剂在两种人肝癌细胞模型中的比较。

RNA干扰是了解基因功能的强大方法，可用于治疗和实验目的。由于缺乏对各种肝细胞系基因沉默的知识，因此本工作旨在在两种人类细胞模型中比较两种转染剂，即基于脂质体的Lipofectamine™RNAiMAX和基于HepG2的，基于聚合物的GenMute™。肝癌，HepG2和Huh7.5。在第一部分中，我们通过细胞成像分析评估了荧光Cy3标记的阴性对照siRNA的转染效率。我们发现，与Lipofectamine RNAiMAX转染的细胞相比，用GenMute处理的细胞在HepG2和Huh7.5细胞中均表现出较高的荧光阴性对照siRNA摄取。在第二部分中 我们在每次转染处理后通过RT-PCR评估了两种转染试剂对GAPDH沉默的相似GAPDH mRNA表达。最后，我们通过MTT测定法测量了细胞活力，观察到用GenMute转染的细胞相对于Lipofectamine RNAiMAX给药的细胞具有更高的活力。这些结果表明GenMute试剂可能被认为是最适合肝基因沉默的转染剂。