Back to Journals » International Journal of Nanomedicine » Volume 6

Uptake and intracellular traffic of siRNA dendriplexes in glioblastoma cells and macrophages

Authors Perez, Cosaka, Romero EL, Morilla MJ

Published 4 November 2011 Volume 2011:6 Pages 2715—2728


Review by Single-blind

Peer reviewer comments 2

Ana Paula Perez, Maria Luz Cosaka, Eder Lilia Romero, Maria Jose Morilla
Programa de Nanomedicinas, Departamento de Ciencia y Tecnología, Universidad Nacional de Quilmes, Bernal, Buenos Aires, Argentina

Background: Gene silencing using small interfering RNA (siRNA) is a promising new therapeutic approach for glioblastoma. The endocytic uptake and delivery of siRNA to intracellular compartments could be enhanced by complexation with polyamidoamine dendrimers. In the present work, the uptake mechanisms and intracellular traffic of siRNA/generation 7 dendrimer complexes (siRNA dendriplexes) were screened in T98G glioblastoma and J774 macrophages.
Methods: The effect of a set of chemical inhibitors of endocytosis on the uptake and silencing capacity of dendriplexes was determined by flow cytometry. Colocalization of fluorescent dendriplexes with endocytic markers and occurrence of intracellular dissociation were assessed by confocal laser scanning microscopy.
Results: Uptake of siRNA dendriplexes by T98G cells was reduced by methyl-ß-cyclodextrin, and genistein, and cytochalasine D, silencing activity was reduced by genistein; dendriplexes colocalized with cholera toxin subunit B. Therefore, caveolin-dependent endocytosis was involved both in the uptake and silencing activity of siRNA dendriplexes. On the other hand, uptake of siRNA dendriplexes by J774 cells was reduced by methyl-ß-cyclodextrin, genistein, chlorpromazine, chloroquine, cytochalasine D, and nocodazole, the silencing activity was not affected by chlorpromazine, genistein or chloroquine, and dendriplexes colocalized with transferrin and cholera toxin subunit B. Thus, both clathrin-dependent and caveolin-dependent endocytosis mediated the uptake and silencing activity of the siRNA dendriplexes. SiRNA dendriplexes were internalized at higher rates by T98G but induced lower silencing than in J774 cells. SiRNA dendriplexes showed relatively slow dissociation kinetics, and their escape towards the cytosol was not mediated by acidification independently of the uptake pathway.
Conclusion: The extent of cellular uptake of siRNA dendriplexes was inversely related to their silencing activity. The higher silencing activity of siRNA dendriplexes in J774 cells could be ascribed to the contribution of clathrin-dependent and caveolin-dependent endocytosis vs only caveolin-dependent endocytosis in T98G cells.

Keywords: silencing, dendrimers, clathrin, caveolin

Creative Commons License This work is published and licensed by Dove Medical Press Limited. The full terms of this license are available at and incorporate the Creative Commons Attribution - Non Commercial (unported, v3.0) License. By accessing the work you hereby accept the Terms. Non-commercial uses of the work are permitted without any further permission from Dove Medical Press Limited, provided the work is properly attributed. For permission for commercial use of this work, please see paragraphs 4.2 and 5 of our Terms.

Download Article [PDF]  View Full Text [HTML][Machine readable]


Other articles by this author:

Selective cytotoxicity of PAMAM G5 core–PAMAM G2.5 shell tecto-dendrimers on melanoma cells

Schilrreff P, Mundiña-Weilenmann C, Romero EL, Morilla MJ

International Journal of Nanomedicine 2012, 7:4121-4133

Published Date: 31 July 2012

Increased brain radioactivity by intranasal 32P-labeled siRNA dendriplexes within in situ-forming mucoadhesive gels

Perez AP, Mundiña-Weilenmann C, Romero EL, Morilla MJ

International Journal of Nanomedicine 2012, 7:1373-1385

Published Date: 12 March 2012

Readers of this article also read:

Bone repair by periodontal ligament stem cell-seeded nanohydroxyapatite-chitosan scaffold

Ge S, Zhao N, Wang L, Yu M, Liu H, Song A, Huang J, Wang G, Yang P

International Journal of Nanomedicine 2012, 7:5405-5414

Published Date: 10 October 2012

Reduction of atherosclerotic lesions in rabbits treated with etoposide associated with cholesterol-rich nanoemulsions

Tavares ER, Freitas FR, Diament JD, Maranhão RC

International Journal of Nanomedicine 2011, 6:2297-2304

Published Date: 12 October 2011

Doxorubicin-loaded PEG-PCL copolymer micelles enhance cytotoxicity and intracellular accumulation of doxorubicin in adriamycin-resistant tumor cells

Diao YY, Li HY, Fu YH, Han M, Hu YL, Jiang HL, Tsutsumi Y, Wei QC, Chen DW, Gao JQ

International Journal of Nanomedicine 2011, 6:1955-1962

Published Date: 12 September 2011

Bactericidal activities of woven cotton and nonwoven polypropylene fabrics coated with hydroxyapatite-binding silver/titanium dioxide ceramic nanocomposite "Earth-plus"

Kasuga E, Kawakami Y, Matsumoto T, Hidaka E, Oana K, Ogiwara N, Yamaki D, Sakurada T, Honda  T

International Journal of Nanomedicine 2011, 6:1937-1943

Published Date: 9 September 2011

New tumor-targeted nanosized delivery carrier for oligonucleotides: characteristics in vitro and in vivo

Zhou T, Jia X, Li H, Wang J, Zhang H, A Y, Zhang ZZ

International Journal of Nanomedicine 2011, 6:1527-1534

Published Date: 22 July 2011

Poly (ε-caprolactone) nanofibrous ring surrounding a polyvinyl alcohol hydrogel for the development of a biocompatible two-part artificial cornea

Bakhshandeh H, Soleimani M, Shah Hosseini S, Hashemi H, Shabani I, Shafiee A, Behesht Nejad AH, Erfan M, Dinarvand R, Atyabi F

International Journal of Nanomedicine 2011, 6:1509-1515

Published Date: 14 July 2011

Study of pharmacokinetics and tissue distribution of liposomal brucine for dermal administration

Yang B-C, Chu Z-F, Zhu S, Wang L-J, Feng Y-H, Li F-H, Liu C-S, Yuan Y

International Journal of Nanomedicine 2011, 6:1109-1116

Published Date: 26 May 2011

Dispersion stability and exothermic properties of DNA-functionalized single-walled carbon nanotubes

Kawaguchi M, Ohno J, Irie A, Fukushima T, Yamazaki J, Nakashima N

International Journal of Nanomedicine 2011, 6:729-736

Published Date: 8 April 2011