Lectin enhancement of the lipofection efficiency in human lung carcinoma cells.
Lectin enhancement of the lipofection efficiency in human lung carcinoma cells.
复制标题
凝集素增强人肺癌细胞的脂转染效率。
DOI：
10.1016/s0304-4165(99)00100-2
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发表时间：
1999
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,PW
脂质体转染人肺癌细胞的效率低，需要进一步开发更有效的基因递送策略。本研究的目的是确定凝集素是否可以提高肺癌细胞的脂质转染效率。使用脂质体加凝集素作为载体，用含有β-半乳糖苷酶报告基因（pCMVlacZ）的质粒DNA转染生长至90%汇合的A549、Calu 3和H292细胞18 h。十种不同的凝集素，表现出广泛的碳水化合物结合特异性，检查其能力，以提高效率的脂转染。通过β-半乳糖苷酶活性（单位/μg蛋白质）和X-Gal染色后的蓝色细胞%评估转染细胞的转染效率。补充有Griffonia simplicifolia-I（GS-I）的Lipofectin在A549和Calu 3细胞中产生最大的脂质转染效率增强（分别为5.3倍和28倍）。怀槐在H292细胞中的脂质转染效率得到最大的增强（6.5倍）。转染效率与递送到细胞核的DNA的量相关。α-甲基-d-吡喃半乳糖苷可抑制FITC标记的GS-Ⅰ的结合和GS-Ⅰ对脂质体转染效率的增强作用，表明α-半乳糖苷介导的基因转移到肺癌细胞中。我们的结论是，凝集素促进脂质体转染是一种有效的基因传递策略。细胞类型特异性凝集素的使用可以允许有效的细胞类型特异性基因靶向。
Poor transfection efficiency of human lung carcinoma cells by lipofection begs further development of more efficient gene delivery strategies. The purpose of this study was to determine whether lectins can improve the lipofection efficiency in lung carcinoma cells. A549, Calu3, and H292 cells grown to 90% confluence were transfected for 18 h with a plasmid DNA containing a β-galactosidase reporter gene (pCMVlacZ) using lipofectin plus a lectin as the vector. Ten different lectins, which exhibit a wide range of carbohydrate-binding specificities, were examined for their abilities to enhance the efficiency of lipofection. The transfected cells were assessed for transfection efficiency by β-galactosidase activity (units/μg protein) and % blue cells following X-Gal stain. Lipofectin supplemented with Griffonia simplicifolia-I (GS-I) yields largest enhancement of the lipofection efficiency in A549 and Calu3 cells (5.3- and 28-fold, respectively). Maackia amurensis gives the largest enhancement (6.5-fold) of lipofection efficiency in H292 cells. The transfection efficiency correlates with the amounts of DNA delivered to the nucleus. Binding of FITC-labeled GS-I and the enhancement of the lipofection efficiency by GS-I were inhibited by α-methyl-d-galactopyranoside, indicating an α-galactoside-mediated gene transfer to lung carcinoma cells. We conclude that lectin-facilitated lipofection is an efficient gene delivery strategy. Employment of cell type-specific lectins may allow for efficient cell type-specific gene targeting.