Transmembrane routes of cationic liposome-mediated gene delivery using human throat epidermis cancer cells |
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Authors: | Shaohui Cui Bing Wang Yinan Zhao Huiying Chen Huiqin Ding Defu Zhi Shubiao Zhang |
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Institution: | 1. SEAC-ME Key Laboratory of Biotechnology and Bio-resouces Utilization, College of Life Science, Dalian Nationalities University, Dalian, 116600, Liaoning, China
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Abstract: | 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). |
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