Ultrasound enhances liposome-mediated gene transfection
Ultrasound enhances liposome-mediated gene transfection
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DOI：
10.1016/j.ultsonch.2004.06.006
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发表时间：
2005-08-01
影响因子：
8.4
通讯作者：
Kondo, T
中科院分区：
文献类型：
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作者：
Feril, LB;Ogawa, R;Kondo, T
以往的研究表明，一些系列的脂质体，通常含有阳离子脂质，是有用的工具，基因导入细胞。为了研究超声（US）对脂质体介导的转染的影响，在本研究中使用了三种类型的脂质体（以增加转染效率的顺序命名为L1、L2和L3），其含有不同比例的O，O '-双十四酰基-N-（α-三甲基铵乙酰基）二乙醇胺氯化物、二油酰基磷脂酰乙醇胺和/或胆固醇。在超声处理之前，用含有荧光素酶基因的脂质体-DNA复合物处理HeLa细胞2小时。增强的最佳US条件被确定为0.5 W/cm（2），1 MHz连续波1 min，并且基于自由基的EPR检测高于惯性空化阈值。处理后24小时，荧光素酶表达通过超声处理显著增加，L1为2.4倍，L2为1.7倍。然而，对于在脂质体中表现出最高表达水平的L3，当DNA-L3处理后15分钟进行超声处理时观察到显著但最小的增强，这表明脂质体的效率也决定了超声处理的适当时机。对于L1和L2，需要用脂质体-DNA复合物预超声孵育2小时（对于L3，30分钟）以获得增强，这表明US仅在细胞具有足够的DNA摄取后才能增强转染。(c)2004 Elsevier B. V.保留所有权利。
Previous studies have shown that some series of liposomes, usually containing cationic lipids, are useful tools for gene introduction into cells. To investigate the effect of ultrasound (US) on liposome-mediated transfection, three types of liposomes (designated L1, L2 and L3, in the order of increasing transfection efficiency) containing O,O'-ditetradecanoyl-N-(alpha-trimethylammonioacetyl) diethanolamine chloride, dioleoylphosphatidylethanolamine, and/or cholesterol at varying ratios, were used in this study. HeLa cells were treated with liposome-DNA complexes containing luciferase genes for 2 h before sonication. Optimal US condition for the enhancement was determined to be 0.5 W/cm(2), 1 MHz continuous wave for 1 min and was above threshold for inertial cavitation based on EPR detection of free radicals. Luciferase expressions 24 h after the treatments were significantly increased by sonication to 2.4 fold with L1, and 1.7 fold with L2. However, with L3, which showed the highest level of expression among the liposomes, significant but minimal enhancement was observed when sonication was done 15 min after the DNA-L3 treatment, suggesting that efficiency of the liposome also determines the proper timing for sonication. The 2 h pre-sonication incubation with liposome-DNA complexes for L1 and L2 (30 min for L3) required to attain enhancement, suggests that US works to enhance transfection only after cells had enough DNA uptake. (c) 2004 Elsevier B.V. All rights reserved.