Biotechnology Letters, 2014, 36(1):1-7.

Transmembrane routes of cationic liposome-mediated gene delivery using human throat epidermis cancer cells

Shaohui Cui, Bing Wang, Yinan Zhao, et al

SEAC-ME Key Laboratory of Biotechnology and Bio-resouces Utilization, College of Life Science, Dalian Nationalities University, Dalian, 116600, Liaoning, China

For studying the mechanism of cationic liposome-mediated transmembrane routes for gene delivery, various inhibitors of endocytosis were used to treat human throat epidermis cancer cells, Hep-2, before transfection with Lipofectamine 2000/pGFP-N2 or Lipofectamine 2000/pGL3. To eliminate the effect of inhibitor toxicity on transfection, the RLU/survival rate was used to represent the transfection efficiency. Chlorpromazine and wortmannin, clathrin inhibitors, decreased transfection efficiency by 44 % (100 μM) and 31 % (100 nM), respectively. At the same time, genistein, a caveolin inhibitor, decreased it by 30 % (200 μM). Thus combined transmembrane routes through the clathrin and caveolae-mediated pathways were major mechanisms of cell uptake for the cationic liposome-mediated gene delivery. After entering the cells, microtubules played an important role on gene delivery as vinblastine, a microtubulin inhibitor, could reduce transfection efficiency by 41 % (200 nM).

荷兰《生物技术快讯》，2014年1月

使用人体咽喉表皮肿瘤细胞进行阳离子脂质体介导的基因传递的跨膜路线

崔绍辉、 王兵、 赵一楠、等

中国辽宁大连民族大学石生命科学学院生物技术和生物资源利用SEAC-ME重点实验室

为了研究阳离子脂质体介导的基因传递的跨膜路线机制，在脂质体 2000/pGFP-N2 或脂质体 2000/pGL3 转染之前，多种细胞内吞作用的抑制剂被用于治疗人体喉咙表皮肿瘤细胞，Hep-2。为了消除抑制剂毒性对转染的影响，RLU/成活率的比值被用来表示转染效率。氯普鲁马嗪和渥曼青霉素，内涵蛋白的抑制剂，分别降低了44% (100 μ M) 和 31%(100 nM)的 转染效率。同时，金雀异黄素，一种小窝蛋白抑制剂，降低了 30%(200 μM)。因此通过内涵蛋白和小窝蛋白调节的通路联合的跨膜路线是为了阳离子脂质体介导的基因传递的细胞吸收的主要机制。在进入细胞后，微管蛋白对基因传递发挥重要作用，例如长春碱，一种微管蛋白抑制剂，可以降低41%的转染效率 (200 nM)。