Genes & CellsGenes & Cells2313-18292500-2562Human Stem Cells Institute12158310.23868/gc121583Gene transfer using new complexes between cardiolipin-like dicationic lipids and plasmid DNA to tumor cellsZhdanovR IKazan (Volga region) Federal University, Kazan;Institute General Pathology and Pathophysiology of RAMS, Moscow;-MoskovtsevA AInstitute General Pathology and Pathophysiology of RAMS, Moscow;Blokhin Russian Cancer Research Center of RAMS, Moscow;-BlokhinD YuBlokhin Russian Cancer Research Center of RAMS, Moscow-DoynikovaA NКazan (Volga region) Federal University, Kazan-ShakirovaR IКazan (Volga region) Federal University, Kazan-SebyakinYu LMoscow State University of Fine Chemical Technology, Moscow-Kazan (Volga region) Federal University, KazanInstitute General Pathology and Pathophysiology of RAMS, MoscowBlokhin Russian Cancer Research Center of RAMS, MoscowКazan (Volga region) Federal University, KazanMoscow State University of Fine Chemical Technology, Moscow1509201273576111012023Copyright © 2012, Eco-Vector2012The lipid vesicles of bisamphiphiles cardiolipin-like dicationic
lipids (CDL) I-IV were studied for creation of lipoplexes with
plasmid DNA of different sizes to obtain stable lipoplexes for
gene transfer to gene therapy. Lipoplexes sizes (300±100 nm)
and stablity (> 2 hrs) of CDL were sufficient to be used in
gene transfer against monolayer and suspension cell cultures.
The CDL total cytotoxicity determined by MTT-test was lower
compare to lipofectin as a control. Transfection conditions
against tumor cells lines were optimized by lipoplexes of CDL
and plasmid DNA. The most efficient transfection for lipoplexes
CDL-plasmid DNA was at the lipid-DNA (L/D) ratio equal to 5
(for lipofectin, it was 2). For monolayer cell cultures, lipoplexes
CDL-I are comparable in terms of transfection efficacy with
lipofectin; in the case of suspension culture, their efficiency
was lower by one order of magnitude. It permits a usage
of lipoplexes suggested as mediators for gene transfer and
delivery to human tumor cells.dicationic lipidslipofectionreporter genegene transfercell cultureдикатионные липидылипофекциярепортерные геныгенный переноскультуры клеток[Felgner P.L., Gadek T.R., Holm M. et al. Lipofectin: a highly efficient lipid-mediated DNA transfection procedure. PNAS USA 1987; 84: 7413-7.][Zhdanov R.I., Bogdanenko E.V., Moskovtsev A.A. et al. Liposomemediated gene delivery: dependence on lipid structure, glycolipid mediated targeting, and immunological properties. In: Duzgunes N., editor. Methods in Enzymol. Liposomes. Part C: Gene Transfer and Therapy: Elsevier. 2003; 373: 433-65.][Жданов Р.И., Хусаинова Р.С., Иваницкий Г.Р. и др. Невирус- ные векторы в генной терапии. Новый подход в липофекции. Вопро- сы Биол. Мед. Фарм. Химии. 2000; 1: 10-17.][Duzgunes N. de Ilarduya C., Simoes S. et al. Cationic liposomes for gene delivery: novel cationic lipids and enhancement by proteins and peptides. Curr. Med. Chem. 2003; 10(14): 1213-20.][Zhdanov R.I., Podobed O.V., Vlassov V.V. Cationic lipid-DNA complexes - lipoplexes-for gene transfer and therapy. Bioelectrochem. 2002; 58(1): 53-64.][Akinc A., Anderson D.G., Lynn D.M. et al. Synthesis of poly(betaamino ester)s optimized for highly effective gene delivery. Bioconjug. Chem. 2003; 14(5): 979-88.][Zabner J., Fasbender A.J., Moninger T. et al. Cellular and molecular barriers to gene transfer by a cationic lipids. J. Biol. Chem. 1995; 270(18): 18997-9007.][Wrobel I., Collins D. Fusion of cationic liposomes with mammalian cells occurs after endocytosis. Biochim Biophys Acta. 1995; 1235(2): 296-304.][Lasic D.D. Liposomes in gene delivery. NY: CRC Press, Boca Raton; 1997.][Leong K.W., Mao H.Q., Truong-Le V.L. et al. DNA-polycation nanospheres as non-viral gene delivery vehicles. J. Control. Release. 1998; 53(1-3): 183-93.][Ravi Kumar M.N., Bakowsky U., Lehr C.M. Preparation and characterization of cationic PLGA nanospheres as DNA carriers. Biomaterials 2004; 25(10): 1771-7.][Templeton N.S., Lasic D.D., Frederik P.M. et al. Improved DNA: liposome complexes for increased systemic delivery and gene expression. Nature Biotechnology 1997; 15: 647-52.][Menger F.M., Keiper J.S. Gemini surfactants. Angew. Chem. Int. Ed. 2000; 39: 1906-20.][Hattori T., Adachi K., Shizuri Y. New ceramide from marine sponge Haliclona koremella and related compounds as antifouling substances against macroalgae. J. Nat. Prod. 1998; 61: 823-6.][Полякова А.А., Панченкова И.А., Скрипникова М.А. и др. Мембранообразующие свойства димерных катионных амфифи- лов на основе L-глутаминовой кислоты. Биологические мембраны 2003; 20(2): 178-83.][Московцев А.А. Индукция гибели опухолевых клеток чело- века новой невирусной системой на основе дикатионных липидов, гена тимидинкиназы HVS-tk и ганцикловира [диссертация]. Москва: НИИОПП РАМН; 2007.][Соколовская А.А., Заботина Т.Н., Блохин Д.Ю. и др. СD95-дефицитные клетки сублинии Jurkat/A4, устойчивые к лекарственно-индуцированному апоптозу. Экспер. Онкология 2001; 23: 175-80.][Маниатис Т., Фрич Э., Сэмбрук Дж. Молекулярное клониро- вание Методы генетической инженерии. Москва: Мир; 1984.][Mozafari M.R., Flanagan J., Zhu X. et al. Sample protocols for the preparation and characterisation of lipid vesicles In: Mozafari][M.R., Mortazavi S.M., editors. Nanoliposomes: From fundamentals to recent developments. Oxford: Trafford Publishing Ltd; 2005. p. 125-45.][Niks M., Otto M. Towards an optimized MTT assay. J. Immunol. Meth. 1990; 130(1): 149-51.][Жданов Р.И., ЭльКади А., Московцев А.А. и др. Зависи- мость активности генного переноса липоплексами на основе но- вых дикатионных липидов от их строения. Докл. Акад. наук 2004; 398(1): 118-21.]