Protein Expression Knockdown in Cancer Cells Induced by a Gemini Cationic Lipid Nanovector with Histidine-Based Polar Heads

Pharmaceutics

Pablo Taboada¹  EvaM. Villar¹  María Martínez-Negro²  Andrés Guerrero-Martínez²  Elena Junquera²  Natalia Sánchez-Arribas²  Emilio Aicart²  José Osío Barcina³  Lourdes Pérez⁴ 

[1] Departamento de Física de Partículas, Facultad de Físicas e Instituto de Investigaciones Sanitarias (IDIS), Universidad de Santiago de Compostela, Campus Vida, E-15782 Santiago de Compostela, Spain;Departamento de Química Física, Facultad de Ciencias Químicas, Universidad Complutense de Madrid, 28040 Madrid, Spain;Departamento de Química Orgánica, Facultad de Ciencias Químicas, Universidad Complutense de Madrid, 28040 Madrid, Spain;Departamento de Tensioactivos y Nanobiotecnología, IQAC-CSIC, 08034 Barcelona, Spain;

关键词: Non-viral gene delivery;    gene knockdown efficiency;    small interfering RNA;    amino acid-based gemini cationic lipids;    protein expression;    protein corona;   

DOI  :  10.3390/pharmaceutics12090791

来源: DOAJ

【 摘 要 】

A histidine-based gemini cationic lipid, which had already demonstrated its efficiency as a plasmid DNA (pDNA) nanocarrier, has been used in this work to transfect a small interfering RNA (siRNA) into cancer cells. In combination with the helper lipid monoolein glycerol (MOG), the cationic lipid was used as an antiGFP-siRNA nanovector in a multidisciplinary study. Initially, a biophysical characterization by zeta potential (ζ) and agarose gel electrophoresis experiments was performed to determine the lipid effective charge and confirm siRNA compaction. The lipoplexes formed were arranged in L_α lamellar lyotropic liquid crystal phases with a cluster-type morphology, as cryo-transmission electron microscopy (cryo-TEM) and small-angle X-ray scattering (SAXS) studies revealed. Additionally, in vitro experiments confirmed the high gene knockdown efficiency of the lipid-based nanovehicle as detected by flow cytometry (FC) and epifluorescence microscopy, even better than that of Lipofectamine2000*, the transfecting reagent commonly used as a positive control. Cytotoxicity assays indicated that the nanovector is non-toxic to cells. Finally, using nano-liquid chromatography tandem mass spectrometry (nanoLC-MS/MS), apolipoprotein A-I and A-II followed by serum albumin were identified as the proteins with higher affinity for the surface of the lipoplexes. This fact could be beyond the remarkable silencing activity of the histidine-based lipid nanocarrier herein presented.

【 授权许可】

Unknown