阳离子脂质体介导RNA干扰的效果评价

Efficacy of RNA interference mediated by cationic liposomes

摘要:

考察自制的肽型阳离子脂质体CDO14作为RNA转染载体的细胞毒性及其运载siRNA进行RNA干扰的效果。通过MTT法检测脂质体对稳定表达荧光素酶的肺癌A549 (Luc-A549) 细胞的毒性。以脂质体为载体将荧光素酶siRNA (Luc-siRNA) 转染至Luc-A549细胞内，用发光仪检测转染细胞内荧光素酶含量，BCA法检测细胞内总蛋白含量。在裸鼠腋下接种Luc-A549细胞，成瘤后尾静脉注射Luc-siRNA和脂质体的复合物，利用活体成像系统检测模型小鼠体内荧光素酶的表达量。细胞毒性实验表明，自制脂质体的毒性与商品脂质体DOTAP相近，低于商品脂质体Lipo2000；细胞转染实验表明自制脂质体作为基因转染载体的转染效率高于DOTAP；体内转染实验表明CDO14作为载体转染效果优于DOTAP。结果表明，肽型阳离子脂质体CDO14具有毒性小、转染效率高等优点，有望作为转染载体用于基因治疗。

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.