PEG-G5.NH2-FITC-DOTA(Gd)-Monalizumab/IPH4301 探针的初步研究.

Objective: To synthesize PEG-G5.NH2-FITC-DOTA (Gd) -Monalizumab/IPH4301 nanoprobes, and investigate the ability of PEG-G5.NH2-FITC-DOTA (Gd) -Monalizumab/IPH4301 nanoprobe bound to NK-92MI cells, effect on cell viability, magnetic resonance imaging in vitro and promotion of the apoptosis of targeted tumor cells by NK-92MI cells. Methods: PEG-G5.NH2-FITC-DOTA (Gd) -Monalizumab/IPH4301 nanoprobes were synthesized and characterized by Transmission Electron Microscope (TEM) and its particle size was calculated using Image J software. Furthermore, flow cytometry was used to analyze the cellular uptake of nanoprobes to NK-92MI cells and magnetic resonance imaging was applied to measure the T1 signal of NK-92MI cells treated with different concentration of nanoprobes. Finally, the expressions of apoptosis-related proteins, such as Cleaved-caspase3, Bax, Bcl-2, were investigated by Western Blot in one of triple-negative breast cancer cell line (MDA-MB-231) after co-culture with NK-92MI cells at ratio of 1:5 overnight and the concentrations of interferon-γ were detected by Elisa assay in the medium. Results: The prepared nanoprobes had uniform particle size distribution and the morphology was nearly spherical. The amounts of nanoprobes bound to NK-92MI cells and T1 signal detected by MRI gradually increased with rising concentration. Moreover, nanoprobes modified with Monalizumab and IPH4301 antibodies promoted the release of interferon-γ in the medium and the apoptosis of MDA-MB-231 cells after cocultured with NK-92MI cells overnight. Conclusions: After binding to NK-92MI cells, the PEG-G5.NH2-FITC-DOTA (Gd) -Monalizumab/IPH4301nanoprobes could be visualized with 3.0T MRI in vitro and enhanced cytotoxic effect of NK-92MI cells to induce the apoptosis of MDA-MB-231 cells. [ABSTRACT FROM AUTHOR]