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阳离子脂质体介导的RNA干扰的疗效

[Efficacy of RNA interference mediated by cationic liposomes].

作者信息

Han Wenqi, Zhen Yuhong, Zhang Shubiao, Zhao Yinan, Sun Yong, Guo Xin, Wang Enxia, Liu Zi, Sun Yaoting

出版信息

Sheng Wu Gong Cheng Xue Bao. 2015 Aug;31(8):1239-46.

Abstract

To investigate the cytotoxicity of the homemade peptide cationic liposome CDO14 and its efficacy of RNA interference (RNAi). MTT method was used to determine the cytotoxicity of the liposome to a human lung cancer cell line Luc-A549 that can express luciferase stably. Luciferase siRNA (Luc-siRNA) was transfected into Luc-A549 cells by CDO14. Contents of luciferase in the transfected cells were detected by luminous instrument and contents of total protein in these cells were detected by BCA method. Nude mice were inoculated with Luc-A549 cells in axilla to establish xenograft tumor model. Complexes of Luc-siRNA and the cationic liposomes were injected into the modeling mice via tail vein. Contents of luciferase in the transfected mice were detected by the whole body imaging system. The cytotoxicity of the homemade cationic liposome was similar to that of commercial liposome DOTAP, and lower than that of Lipo2000. The siRNA transfection efficacy mediated by CDO14 was higher than that mediated by DOTAP. The homemade peptide cationic liposome CDO14 is expected to serve as delivery vector in gene therapy because of its low cytotoxicity and high transfection efficiency.

摘要

研究自制肽阳离子脂质体CDO14的细胞毒性及其RNA干扰(RNAi)效果。采用MTT法测定脂质体对可稳定表达荧光素酶的人肺癌细胞系Luc-A549的细胞毒性。通过CDO14将荧光素酶siRNA(Luc-siRNA)转染至Luc-A549细胞。用发光仪检测转染细胞中荧光素酶含量,用BCA法检测这些细胞中总蛋白含量。将Luc-A549细胞接种于裸鼠腋窝建立异种移植瘤模型。将Luc-siRNA与阳离子脂质体的复合物经尾静脉注入建模小鼠体内。用全身成像系统检测转染小鼠体内荧光素酶含量。自制阳离子脂质体的细胞毒性与市售脂质体DOTAP相似,低于Lipo2000。CDO14介导的siRNA转染效率高于DOTAP。自制肽阳离子脂质体CDO14因其低细胞毒性和高转染效率有望成为基因治疗中的递送载体。

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