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    Her2/neu small interfering RNA delivered in culture by a streptavidin nanoparticle

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    Authors
    Liu, Xinrong
    Nakamura, Kayoko
    Cheng, Dengfeng
    Peng, Cong
    Xiao, Nan
    Liu, Yuxia
    Chen, Ling
    Rusckowski, Mary
    Hnatowich, Donald J.
    UMass Chan Affiliations
    Department of Radiology, Division of Nuclear Medicine
    Document Type
    Journal Article
    Publication Date
    2012-07-01
    Keywords
    2',5'-Oligoadenylate Synthetase
    Antibodies, Monoclonal
    Antibodies, Monoclonal, Humanized
    Biotinylation
    Cell Line, Tumor
    Drug Delivery Systems
    Female
    *Genes, erbB-2
    Humans
    Interferons
    Nanoparticles
    RNA, Messenger
    RNA, Small Interfering
    Receptor, ErbB-2
    STAT1 Transcription Factor
    Streptavidin
    Transfection
    delivery nanoparticle
    Her2
    siRNA
    streptavidin
    radiolabeled oligomer
    Nanomedicine
    Neoplasms
    Oncology
    Radiology
    Therapeutics
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    Link to Full Text
    http://dx.doi.org/10.2174/156720112801323035
    Abstract
    A three-component nanoparticle consisting of biotinylated Trastuzumab antiHer2 antibody, tat transferring peptide and radiolabeled antisense oligomer, linked together through streptavidin, have shown promise in the delivery to Her2+ tumor in mice following intravenous administration and with evidence of radiotherapeutic efficacy. These results have encouraged us to consider the nanoparticle as a delivery vehicle for RNA interference therapy in which the radiolabeled antisense oligomer is replaced with an unlabeled siRNA duplex. The siRNA stability within the nanoparticle was first confirmed by incubation with RNase A. The interferon responses, that indicate off-target cytotoxicity, were evaluated by quantitative real-time RT-PCR in BT-474 (Her2+) human breast cancer cells by measuring the mRNA expression of 2', 5'-oligoadenylate synthetase (OAS1) and Stat-1, two key interferon-responsive genes. Thereafter the cytotoxicity induced by the siRNA nanoparticle was evaluated by a clonogenic survival assay in BT-474 cells while the Her2 expression of these target cells was evaluated for evidence of specific gene silencing. The siRNA within the three-component anti- Her2/neu siRNA nanoparticle was largely protected from RNase-dependent degradation and did not activate an interferon response. The nanoparticle effectively and significantly inhibited colony formation of the target cells and silenced the Her2 gene expression at 5 nM compared with the identical nanoparticle with a scrambled siRNA. Our delivery nanoparticle, with tumor targeting provided by the antibody and its accumulation without entrapment, possibly due to the transfecting peptide, delivered an siRNA duplex to the proper subcellular localization for specific and effective gene silencing in culture by what appears to be an siRNA mechanism.
    Source
    Curr Drug Deliv. 2012 Jul;9(4):431-6. doi:10.2174/156720112801323035
    DOI
    10.2174/156720112801323035
    Permanent Link to this Item
    http://hdl.handle.net/20.500.14038/48002
    PubMed ID
    22520071
    Related Resources
    Link to Article in PubMed
    ae974a485f413a2113503eed53cd6c53
    10.2174/156720112801323035
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