非病毒载体介导的基因转染效率及其对人舌鳞癌细胞生长增殖的影响

目的]观察多种非病毒载体介导增强型绿色荧光蛋白(EGFP)基因在舌鳞癌细胞内的表达情况,评价各载体的转染效率及其对细胞生长增殖的影响.方法]采用pEGFP-N1评估阳离子脂质体、聚酰胺-胺型树枝状高聚物(PAMAM-D)以及SuperFect纳米微粒在舌鳞癌Tca8113细胞中的转染效率;绘制生长曲线,计算细胞的相对存活率,流式细胞仪检测细胞增殖与凋亡.结果]第5代(G5)PAMAM-D的转染效率明显高于第2代(G2)PAMAM-D(42.1%vs 19.4%,P=0.003),与SuperFect和脂质体转染组的结果无统计学差异(42.1%vs35.9%和42.1%vs47.7%,P＞0.05).G5 PAMAM-D对细胞的生长增殖无明显影响(P＞0.05),而脂质体转染组细胞的生长增殖受到明显的抑制(P=0.001),细胞凋亡水平升高(P=0.01).结论]G5 PAMAM-D载体安全低毒,转染效率高,是一种有良好应用前景的新型纳米载体基因转移系统.

Efficiencies of Several Non-viral Vectors-mediated Gene Transfection and Their Influence on Tongue Squamous Carcinoma Cells In Vitro

Objective] To evaluate the gene transfection efficiencies of several non-viral vectors in tongue carcinoma Tca8113 cells, and to investigate their influence on biological behaviors of the tumor cells. Methods]The transfection efficiencies of polyamidoamine dendrimer (PAMAM-D), SuperFect and cationic Lipofectamine 2000 in Tca8113 cells were evaluated using PEGFP-N1. Moreover, multiple biological behaviors of the tumor cells such as growth and proliferating potential were also determined using growth curve depiction, MTT assay, and flow cytometry. Results] The transfection efficiency of the generation 5 (G5) PAMAM-D group was higher than that of the generation 2 (G2) PAMAM-D group (42.1% vs 19.4%, P=0.003) while had no statistical difference compared to SuperFect or cationic lipids group (42.1% vs 35.9% and 42.1% vs 47.7%, P > 0.05). The growth and proliferation activity of the tumor cells in G5 PAMAM-D group was not suppressed compared to untransfected group (P > 0.05), whereas in cationic lipids group was significantly inhibited in a dose-dependent manner (P < 0.01) and the apoptotic level was increased (P=0.01). Conclusions]G5 PAMAM-D can efficiently transfer reporter gene into Tca8113 cells with no significant cytotoxicity, suggesting its potential as gene delivery vector for tongue carcinoma gene therapy.