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首页> 外文期刊>Asian Pacific Journal of Cancer Prevention >Optomizing Transfection Efficiency of Cervical Cancer Cells Transfected by Cationic Liposomes LipofectamineTM2000
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Optomizing Transfection Efficiency of Cervical Cancer Cells Transfected by Cationic Liposomes LipofectamineTM2000

机译:优化阳离子脂质体脂质膜脂素2000转染的宫颈癌细胞的转染效率

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Background: Currently, cationic liposome has become the commonly used vehicles for gene transfection. Furthermore, one of the most significant steps in microRNAs expression studies is transferring microRNAs into cell cultures successfully. In this study we aim to approach the feasibility of transfection of cervical cancer cell lines mediated by liposome and to obtain the optimized transfection condition for cervical cancer cell lines. Materials and Methods: as the carrier, miR-101 mimic was transfected into Hela cells and Siha cells. Using green fluorescent protein as reporter gene, to set different groups according to cell seeding density, the amount of miRNA, miRNA and the proportion of Liposomes, Whether to add serum into medium to study their impact on the liposomal transfection efficiency. Finally, MTT assay was used to analyze the relative minimal cell toxicity of liposome reagents. Results: The seeding density of Hela cell line and Siha are (per well of 24 well plates), miRNA amount is 1ul of both, the ratio of miRNA and liposome is 1:0.5 of Hela cell line; 1:0.7 of Siha cell line respectively, after 24 hours we can get the highest transfection efficiency. Compared with serum medium, only Siha cells cultured with serum-free medium obtained higher transfection efficiency before transfection (P0.01). MTT assay showed that according to the above conditions which has the lowest cytotoxicity. Conclusions: The method of Liposome to transfected is a suitable way and it can be an efficient reagent for miRNA delivery for Hela cells and Siha cells in vitro. It may serve as a reference for the further research or application.
机译:背景:目前,阳离子脂质体已成为基因转染的常用车辆。此外,MicroRNA表达研究中最重要的步骤之一是将MicroRNA转移到细胞培养物中。在这项研究中,我们的目的是接近脂质体介导的宫颈癌细胞系转染的可行性,并获得宫颈癌细胞系的优化转染条件。材料和方法:作为载体,将miR-101模拟物转染到HeLa细胞和SiHa细胞中。使用绿色荧光蛋白作为报告基因,根据细胞播种密度,miRNA,miRNA和脂质体比例的不同组,是否将血清添加到培养基中,研究它们对脂质体转染效率的影响。最后,使用MTT测定来分析脂质体试剂的相对最小细胞毒性。结果:HeLa细胞系和SiHa的播种密度(每孔24孔板),miRNA量都是两者的1UL,miRNA和脂质体的比例为1:0.5的HeLa细胞; 1:70.7分别在24小时后,我们可以获得最高的转染效率。与血清培养基相比,只有与无血清培养基培养的Siha细胞在转染前获得了更高的转染效率(P <0.01)。 MTT测定显示,根据具有最低细胞毒性的上述条件。结论:转染的脂质体的方法是合适的方式,它可以是用于MiRNA细胞和体外Siha细胞的MiRNA递送的有效试剂。它可以作为进一步研究或应用的参考。

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