Transmembrane routes of cationic liposome-mediated gene delivery using human throat epidermis cancer cells.
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).
SEAC-ME Key Laboratory of Biotechnology and Bio-resouces Utilization, College of Life Science, Dalian Nationalities University, Dalian, 116600, Liaoning, China.
Shaohui
S
Cui
Bing
B
Wang
Yinan
Y
Zhao
Huiying
H
Chen
Huiqin
H
Ding
Defu
D
Zhi
Shubiao
S
Zhang
D002412:Cations / Q000737:chemistry / Q000378:metabolism; D022461:Caveolins / Q000037:antagonists & inhibitors / Q000378:metabolism; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D016503:Drug Delivery Systems; D004705:Endocytosis / Q000187:drug effects / Q000502:physiology*; D018014:Gene Transfer Techniques; D019833:Genistein / Q000494:pharmacology; D006801:Humans; D008081:Liposomes / Q000737:chemistry / Q000378:metabolism*; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008870:Microtubules / Q000187:drug effects; D010610:Pharyngeal Neoplasms; D014162:Transfection; D014747:Vinblastine / Q000494:pharmacology
D002412:Cations; D022461:Caveolins; D008081:Liposomes; D014747:Vinblastine; D019833:Genistein; D008156:Luciferases