Epidermal growth factor receptor targeted nuclear delivery and high-resolution whole cell X-ray imaging of Fe3O4@TiO2 nanoparticles in cancer cells
| Autor: | Ye Yuan, Caroline Doty, William Liu, Sophie Charlotte Gleber, Si Chen, Claus Flachenecker, Qiaoling Jin, Junjing Deng, Gayle E. Woloschak, Chris Jacobsen, Rachel Mak, Keith Brister, Stefan Vogt, Barry Lai, Tatjana Paunesku |
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| Rok vydání: | 2013 |
| Předmět: |
Cytoplasm
Materials science media_common.quotation_subject Active Transport Cell Nucleus General Physics and Astronomy Metal Nanoparticles Ligands Ferric Compounds Article Flow cytometry Neoplasms Fluorescence microscope medicine Humans General Materials Science DNA Breaks Double-Stranded Epidermal growth factor receptor Internalization media_common Cell Nucleus Titanium Drug Carriers medicine.diagnostic_test biology X-Rays General Engineering beta Karyopherins Cell biology ErbB Receptors Cancer cell biology.protein Nanoparticles Beta Karyopherins Comet Assay Intracellular HeLa Cells |
| Zdroj: | ACS nano. 7(12) |
| ISSN: | 1936-086X |
| Popis: | Sequestration within the cytoplasm often limits the efficacy of therapeutic nanoparticles that have specific subcellular targets. To allow for both cellular and subcellular nanoparticle delivery, we have created epidermal growth factor receptor (EGFR)-targeted Fe3O4@TiO2 nanoparticles that use the native intracellular trafficking of EGFR to improve internalization and nuclear translocation in EGFR-expressing HeLa cells. While bound to EGFR, these nanoparticles do not interfere with the interaction between EGFR and karyopherin-β, a protein that is critical for the translocation of ligand-bound EGFR to the nucleus. Thus, a portion of the EGFR-targeted nanoparticles taken up by the cells also reaches cell nuclei. We were able to track nanoparticle accumulation in cells by flow cytometry and nanoparticle subcellular distribution by confocal fluorescent microscopy indirectly, using fluorescently labeled nanoparticles. More importantly, we imaged and quantified intracellular nanoparticles directly, by their elemental signatures, using X-ray fluorescence microscopy at the Bionanoprobe, the first instrument of its kind in the world. The Bionanoprobe can focus hard X-rays down to a 30 nm spot size to map the positions of chemical elements tomographically within whole frozen-hydrated cells. Finally, we show that photoactivation of targeted nanoparticles in cell nuclei, dependent on successful EGFR nuclear accumulation, induces significantly more double-stranded DNA breaks than photoactivation of nanoparticles that remain exclusively in the cytoplasm. |
| Databáze: | OpenAIRE |
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