title: Chitosan polyplex nanoparticle vector for miR-145 expression in MCF-7: Optimization by design of experiment.
abstract: miR-145, a tumor suppressor micro RNA (miRNA), is down regulated in cancer and can be introduced as a therapeutic agent in various cancers including breast cancer. In this study, miR-145 plasmid was transfected to MCF-7 cells using chitosan polyplex nanoparticles. The vector was prepared according to an optimized fabricating method determined by response surface analysis and D-optimal design. Effects of chitosan molecular weight (Mw) and polymer amine to DNA phosphate ratio (N/P) as the variables were investigated on size, zeta potential, stability, and transfection efficiency of the polyplex nanoparticles. The results indicated that there is an interaction between effects of Mw and N/P ratio on the size of nanoparticles. Gel retardation assay demonstrated that the stability of the complexes in serum and preparation medium during storage time depends on the formulation variables. Statistical analysis affirmed that in spite of particle size, the variables of N/P ratio, time of incubation, and zeta potential affect the gene transfection. In conclusion, by selecting the perfect formulation prepared through an optimized method, it is possible to achieve a high transfection efficacy for miR-145 as an anticancer biological macromolecule.
authors.affiliation: Department of Pharmaceutics, Faculty of Pharmacy, Tehran University of Medical Sciences, P.O. Box 14155-6451, Tehran, Iran.
authors.forename: Farnaz Sadat Mirzazadeh
authors.initials: FS
authors.lastname: Tekie
authors.affiliation: Department of Pharmaceutics, Faculty of Pharmacy, Tehran University of Medical Sciences, P.O. Box 14155-6451, Tehran, Iran; Nanotechnology Research Centre, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran. Electronic address: atyabifa@tums.ac.ir.
authors.forename: Fatemeh
authors.initials: F
authors.lastname: Atyabi
authors.affiliation: Department of Molecular Biology and Genetic Engineering, Stem Cell Technology Research Center, P.O. Box 14155-3174, Tehran, Iran; Department of Hematology, School of Medical Sciences, Tarbiat Modares University, P.O. Box 14115-111, Tehran, Iran.
authors.forename: Masoud
authors.initials: M
authors.lastname: Soleimani
authors.affiliation: Department of Molecular Biology and Genetic Engineering, Stem Cell Technology Research Center, P.O. Box 14155-3174, Tehran, Iran.
authors.forename: Ehsan
authors.initials: E
authors.lastname: Arefian
authors.affiliation: Department of Hematology, School of Medical Sciences, Tarbiat Modares University, P.O. Box 14115-111, Tehran, Iran.
authors.forename: Amir
authors.initials: A
authors.lastname: Atashi
authors.affiliation: Department of Pharmaceutics, Faculty of Pharmacy, Tehran University of Medical Sciences, P.O. Box 14155-6451, Tehran, Iran.
authors.forename: Melika
authors.initials: M
authors.lastname: Kiani
authors.affiliation: Department of Drug & Food Control, Faculty of Pharmacy, Tehran University of Medical Sciences, P.O. Box 14155-6451, Tehran, Iran.
authors.forename: Mohammad Reza
authors.initials: MR
authors.lastname: Khoshayand
authors.affiliation: Department of Chemical Medicine, Faculty of Pharmacy, Tehran University of Medical Sciences, P.O. Box 14155-6451, Tehran, Iran.
authors.forename: Mohsen
authors.initials: M
authors.lastname: Amini
authors.affiliation: Nanotechnology Research Centre, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran.
authors.forename: Rassoul
authors.initials: R
authors.lastname: Dinarvand
mesh_terms: D016923:Cell Death / Q000187:drug effects; D048271:Chitosan / Q000737:chemistry* / Q000633:toxicity; D004587:Electrophoresis, Agar Gel; D024202:Electrophoretic Mobility Shift Assay; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D061986:MCF-7 Cells; D035683:MicroRNAs / Q000378:metabolism*; D008962:Models, Theoretical; D053758:Nanoparticles / Q000737:chemistry* / Q000648:ultrastructure; D010316:Particle Size; D010957:Plasmids / Q000378:metabolism; D055672:Static Electricity
chemical_list: C541010:MIRN145 microRNA, human; D035683:MicroRNAs; D049452:Green Fluorescent Proteins; D048271:Chitosan
keywords: Chitosan; Design of experiment; miR-145