Effects of epidermal growth factor, transferrin, and insulin on lipofection efficiency in human lung carcinoma cells.
Effects of epidermal growth factor, transferrin, and insulin on lipofection efficiency in human lung carcinoma cells.
复制标题
表皮生长因子、转铁蛋白和胰岛素对人肺癌细胞脂转染效率的影响。
DOI：
10.1038/sj.cgt.7700092
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发表时间：
2000
期刊：
影响因子：
--
通讯作者：
Cheng,PW
中科院分区：
文献类型：
--
作者：
Yanagihara,K;Cheng,H;Cheng,PW
转染效率低是脂质体转染的主要缺点。我们以前表明，除了转铁蛋白（TF）的Lipofectin增强了120倍的HeLa细胞中的报告基因的表达，并实现了接近100%的转染效率。本研究的目的是确定TF和其他配体是否可以提高肺癌细胞的脂质转染效率。使用Lipofectin加TF、胰岛素或表皮生长因子作为载体，用质粒DNA（pCMVlacZ）转染汇合的A549、Calu 3和H292细胞18小时。通过β-半乳糖苷酶活性（光单位/μg蛋白质）和5-溴-4-氯-3-吲哚基β-D-吡喃半乳糖苷染色后蓝色细胞的百分比评估转染细胞的转染效率。在所有三种细胞系中，补充有表皮生长因子的Lipofectin产生了脂质转染效率的最大增强（比单独的Lipofectin高≤ 23倍）。胰岛素显著提高了A549和Calu 3细胞的脂质转染效率，但在H292细胞中没有，而TF在Calu 3和H292细胞中显示出显著的脂质转染效率提高作用，但在A549细胞中没有。转染效率与递送到细胞核的DNA的量以及受体的量密切相关。这些结果表明，采用配体促进的脂质体转染的基因递送策略可以在人肺癌细胞中实现高转染效率。此外，增强受体的表达可能是提高基因打靶效率的一种可能策略。
Poor transfection efficiency is the major drawback of lipofection. We showed previously that addition of transferrin (TF) to Lipofectin enhanced the expression of a reporter gene in HeLa cells by 120-fold and achieved close to 100% transfection efficiency. The purpose of this study was to determine whether TF and other ligands could improve the efficiency of lipofection in lung carcinoma cells. Confluent A549, Calu3, and H292 cells were transfected for 18 hours with a plasmid DNA (pCMVlacZ) using Lipofectin plus TF, insulin, or epidermal growth factor as the vector. The transfected cells were assessed for transfection efficiency by β-galactosidase activity (light units/μg protein) and the percentage of blue cells following 5-bromo-4-chloro-3-indolyl β-D-galactopyranoside staining. Lipofectin supplemented with epidermal growth factor yielded the largest enhancement of lipofection efficiency (≤ 23-fold over that by Lipofectin alone) in all three cell lines. Insulin significantly enhanced the lipofection efficiency in A549 and Calu3 cells but not in H292 cells, whereas TF showed significant lipofection efficiency-enhancing effect in Calu3 and H292 cells but not in A549 cells. The transfection efficiency correlated well with the amounts of DNA delivered to the nucleus as well as the amounts of the receptor. These results indicate that the gene delivery strategy employing ligand-facilitated lipofection can achieve high transfection efficiency in human lung carcinoma cells. In addition, enhancement of the expression of the receptor may be a possible strategy for increasing the efficiency of gene targeting.