Abstract
Development of biocompatible and multifunctional nanoprobes for tumor targeting, imaging, and therapy still remains a great challenge. Herein, gold nanoparticles (AuNPs) were synthesized in the cavity of horse spleen apoferritin protein (HoSAF) and protein surface was labeled with 2-amino-2-deoxy-glucose (2DG) as a cell surface glucose transport protein specific targeting probe to study the feasibility of its usage as a computer tomography (CT) contrast agent with tumor targeting capability through in vitro experiments. 2DG conjugated and gold-loaded apoferritin (Au-HoSAF-2DG) nanoparticles (NPs) showed selective targeting for human breast adenocarcinoma (MCF-7) cells when compared to normal breast (MCF-10A) cells. This AuNP-based imaging agent was found to be non-cytotoxic in a given concentration range with an apoptotic effect upon longer exposure times towards MCF-7 cells, while MCF-10A cells were affected less. This selective cell death would also be useful for further cancer treatments with the ability of X-ray attenuation in in vitro X-ray and computed tomography (CT) imaging.
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Acknowledgements
We would like to thank Arda Büyüksungur for CT imaging and Seçkin Öztürk for HRTEM imaging at METU.
Funding
This study was funded by Necmettin Erbakan University Research Funding Center (BAP-Project No: 191215003).
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Suppl. Fig. 1 SEC absorbance profiles of HoSAF a) before AuNP formation, b) after AuNP formation with 4000 Au/cage. Elution was monitored both at 280 nm (protein) and 525 nm (AuNP). Suppl. Fig. 2 SEC absorbance profile of Au-HoSAF-2DG. Elution was monitored at both 280 nm (protein) and 525 nm (AuNP). Suppl. Fig. 3 Cell viability of MCF-10A (left) and MCF-7 (right) cells after treatment with 2 different concentrations (0.5 (empty column) and 1.0 mg/mL (filled column)) of HoSAF, Au-HoSAF and Au-HoSAF-2DG NPs for 24 h, 48 h and 72 h (top to bottom). Suppl. Fig. 4 The flow cytometry analysis results of control and after Au-HoSAF-2DG NP treatment at concentration of 1.0 mg/mL for 24 h, 48 h and 72 h of a) MCF-7 and b) MCF-10 A cells. Area A1 and A2 (left, bottom and up) reveals viable and dead cell (necrotic), A3 and A4 (right, top and bottom) correspond to cells undergoing early and late apoptosis, respectively Suppl. Fig. 1 SEC absorbance profiles of HoSAF a) before AuNP formation, b) after AuNP formation with 4000 Au/cage. Elution was monitored both at 280 nm (protein) and 525 nm (AuNP). Suppl. Fig. 2 SEC absorbance profile of Au-HoSAF-2DG. Elution was monitored at both 280 nm (protein) and 525 nm (AuNP). Suppl. Fig. 3 Cell viability of MCF-10A (left) and MCF-7 (right) cells after treatment with 2 different concentrations (0.5 (empty column) and 1.0 mg/mL (filled column)) of HoSAF, Au-HoSAF and Au-HoSAF-2DG NPs for 24 h, 48 h and 72 h (top to bottom). Suppl. Fig. 4 The flow cytometry analysis results of control and after Au-HoSAF-2DG NP treatment at concentration of 1.0 mg/mL for 24 h, 48 h and 72 h of a) MCF-7 and b) MCF-10 A cells. Area A1 and A2 (left, bottom and up) reveals viable and dead cell (necrotic), A3 and A4 (right, top and bottom) correspond to cells undergoing early and late apoptosis, respectively (PDF 686 kb)
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Aslan, T.N., Aşık, E., Güray, N.T. et al. The potential application of gold-apoferritin nanocages conjugated with 2-amino-2-deoxy-glucose for imaging of breast cancer cells. J Biol Inorg Chem 25, 1139–1152 (2020). https://doi.org/10.1007/s00775-020-01830-y
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DOI: https://doi.org/10.1007/s00775-020-01830-y