title: Efficient down-regulation of PKC-α gene expression in A549 lung cancer cells mediated by antisense oligodeoxynucleotides in dendrosomes.
abstract: The completion of human genome project has increased our knowledge of the molecular mechanisms of many diseases, including cancer, thus providing new opportunities for gene therapy. Antisense oligodeoxynucleotides (AsODN) possess great potential as sequence-specific therapeutic agents, which in contrast to classic treatments provide more efficient and target-specific approach to modulate disease-related genes. To be therapeutically effective, sufficient concentrations of intact AsODN must bypass membrane barriers and access the site of action. In this study, a dendrosome delivery strategy was designed to improve the encapsulation of AsODN in non-cationic liposomes to target PKC-α in lung cancer cells in vitro. Subcellular trafficking of fluorescently labeled AsODN was visualized using confocal microscopy. Uptake and expression of mRNA and target protein after AsODN delivery was measured by flow cytometry, qRT-PCR and Western blot analysis, respectively. Dendrosomes showed favorable physicochemical parameters: high encapsulation efficiency and uptake in serum-containing medium with no apparent cytotoxicity. AsODN encapsulated in dendrosome efficiently and specifically suppress the target gene at both mRNA and protein levels. Additional in vivo studies on the application of dendrosome as a delivery system for nucleic acid molecules may lead to improvement of this technology and facilitate the development of therapeutic antisense techniques.
authors.affiliation: Department of Pharmaceutics, School of Pharmacy, Shahid Beheshti University of Medical Sciences, Valiasr Ave., Niayesh Junction, PO Box 14155-6153, Tehran, Iran.
authors.forename: Sara
authors.initials: S
authors.lastname: Movassaghian
authors.forename: Hamid R
authors.initials: HR
authors.lastname: Moghimi
authors.forename: Farshad H
authors.initials: FH
authors.lastname: Shirazi
authors.forename: Alexander
authors.initials: A
authors.lastname: Koshkaryev
authors.forename: Malav S
authors.initials: MS
authors.lastname: Trivedi
authors.forename: Vladimir P
authors.initials: VP
authors.lastname: Torchilin
mesh_terms: D015153:Blotting, Western; D045744:Cell Line, Tumor; D050091:Dendrimers / Q000008:administration & dosage; D015536:Down-Regulation; D005434:Flow Cytometry; D015972:Gene Expression Regulation, Neoplastic; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods; D006801:Humans; D008081:Liposomes; D008175:Lung Neoplasms / Q000235:genetics* / Q000628:therapy; D018613:Microscopy, Confocal; D020319:Oligodeoxyribonucleotides, Antisense / Q000008:administration & dosage*; D051571:Protein Kinase C-alpha / Q000235:genetics*; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D050091:Dendrimers; D008081:Liposomes; D020319:Oligodeoxyribonucleotides, Antisense; D012333:RNA, Messenger; D051571:Protein Kinase C-alpha