Department of Molecular Pharmaceutics, Hoshi University, Tokyo 142-8501, Japan.
Department of Molecular Pathology, Faculty of Pharmaceutical Sciences, Doshisha Women's College of Liberal Arts, Kyotanabe, Kyoto 610-0395, Japan.
Mol Med Rep. 2023 Oct;28(4). doi: 10.3892/mmr.2023.13067. Epub 2023 Aug 18.
Previously, using three types of cationic lipids, the effect of phospholipids in liposomal formulations on gene-knockdown efficacy was determined after and transfection with small interfering RNA (siRNA)/cationic liposome complexes (siRNA lipoplexes) containing various cationic lipids and phospholipids. In the present study, six other types of cationic lipids, namely -dimethyl--tetradecyltetradecan-1-aminium bromide, -hexadecyl--dimethylhexadecan-1-aminium bromide (DC-1-16), 2-[bis{2-(tetradecanoyloxy)ethyl}amino]--trimethyl-2-oxoethan-1-aminium chloride (DC-6-14), 1,2-di--octadecenyl-3-trimethylammonium propane chloride (DOTMA), 1,2-distearoyl-3-trimethylammonium-propane chloride (DSTAP) and 1,2-dioleoyl-3-dimethylammonium-propane were selected, and the effect of phospholipids in liposomal formulations containing each cationic lipid on gene-knockdown was evaluated. A total of 30 types of cationic liposomes composed of each cationic lipid with phosphatidylethanolamine containing unsaturated or saturated diacyl chains (C14, C16 or C18) were prepared. Regardless of the type of cationic lipid, the inclusion of 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) in the liposomal formulations resulted in injectable size of siRNA lipoplexes after mixing of siRNA and cationic liposomes. Transfection of their lipoplexes with luciferase (Luc) siRNA into human breast cancer MCF-7-Luc cells stably expressing Luc led to a strong knockdown of Luc. Furthermore, the systemic injection of siRNA lipoplexes composed of DC-1-16, DC-6-14, DOTMA or DSTAP with DOPE resulted in siRNA accumulation in the lungs. Significant gene-knockdown was observed in the lungs of mice following the systemic injection of siRNA lipoplexes containing DC-1-16 and DOPE. Cationic liposomes composed of DC-1-16 and DOPE serve as potential carriers for and siRNA transfection.
先前，使用三种阳离子脂质体，研究了含有不同阳离子脂质体和磷脂的小干扰 RNA(siRNA)/阳离子脂质体复合物(siRNA 脂质体)转染后，脂质体制剂中的磷脂对基因敲低效果的影响。在本研究中，选择了另外六种类型的阳离子脂质体，即二甲基-二(十四烷基)十四烷-1-铵溴化物、二(十六烷基)二甲基十六烷-1-铵溴化物(DC-1-16)、2-[双(十四烷酰氧基)乙基]氨基]-三甲基-2-氧代乙-1-铵氯化物(DC-6-14)、1,2-二(油酰基)-3-三甲基铵丙烷氯化物(DOTMA)、1,2-二硬脂酰基-3-三甲基铵丙烷氯化物(DSTAP)和 1,2-二油酰基-3-二甲基铵丙烷，评估了含有每种阳离子脂质体的脂质体制剂中磷脂对基因敲低的影响。共制备了 30 种由每个阳离子脂质体与含有不饱和或饱和二酰基链的磷脂酰乙醇胺(C14、C16 或 C18)组成的阳离子脂质体。无论阳离子脂质体的类型如何，在 siRNA 和阳离子脂质体混合后，脂质体制剂中包含 1,2-二油酰基-sn-甘油-3-磷酸乙醇胺(DOPE)会导致 siRNA 脂质体的注射尺寸。用荧光素酶(Luc)siRNA 转染稳定表达 Luc 的人乳腺癌 MCF-7-Luc 细胞的这些脂质体的转染导致 Luc 强烈敲低。此外，含有 DOPE 的 DC-1-16、DC-6-14、DOTMA 或 DSTAP 的 siRNA 脂质体的全身注射导致 siRNA 在肺部积聚。在含有 DC-1-16 和 DOPE 的 siRNA 脂质体的全身注射后，观察到小鼠肺部的显著基因敲低。由 DC-1-16 和 DOPE 组成的阳离子脂质体可作为 和 siRNA 转染的潜在载体。
