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Multifunctional core/shell nanoparticles cross-linked polyetherimide-folic acid as efficient Notch-1 siRNA carrier for targeted killing of breast cancer.

Yang H, Li Y, Li T, Xu M, Chen Y, Wu C, Dang X, Liu Y - Sci Rep (2014)

Bottom Line: Our results showed that Fe3O4@SiO2(FITC)/PEI-FA/Notch-1 shRNA nanoparticles are 64 nm in diameter with well dispersed and superparamagnetic.Magnetic resonance (MR) imaging and fluorescence microscopy showed significant preferential uptake of Fe3O4@SiO2(FITC)/PEI-FA/Notch-1 shRNA nanocomplex by MDA-MB-231 cells.Transfected MDA-MB-231 cells exhibited significantly decreased expression of Notch-1, inhibited cell proliferation, and increased cell apoptosis, leading to the killing of MDA-MB-231 cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Biophysics, School of Life Science and Technology, University of Electronic Science and Technology of China, Chengdu 610054, Sichuan, P.R. China.

ABSTRACT
In gene therapy, how genetic therapeutics can be efficiently and safely delivered into target tissues/cells remains a major obstacle to overcome. To address this issue, nanoparticles consisting of non-covalently coupled polyethyleneimine (PEI) and folic acid (FA) to the magnetic and fluorescent core/shell of Fe3O4@SiO2(FITC) was tested for their ability to deliver Notch-1 shRNA. Our results showed that Fe3O4@SiO2(FITC)/PEI-FA/Notch-1 shRNA nanoparticles are 64 nm in diameter with well dispersed and superparamagnetic. These nanoparticles with on significant cytotoxicity are capable of delivering Notch-1 shRNA into human breast cancer MDA-MB-231 cells with high efficiency while effectively protected shRNA from degradation by exogenous DNaseI and nucleases. Magnetic resonance (MR) imaging and fluorescence microscopy showed significant preferential uptake of Fe3O4@SiO2(FITC)/PEI-FA/Notch-1 shRNA nanocomplex by MDA-MB-231 cells. Transfected MDA-MB-231 cells exhibited significantly decreased expression of Notch-1, inhibited cell proliferation, and increased cell apoptosis, leading to the killing of MDA-MB-231 cells. In light of the magnetic targeting capabilities of Fe3O4@SiO2(FITC)/PEI-FA, our results show that by complexing with a second molecular targeting therapeutic, such as Notch-1 shRNA in this report, Fe3O4@SiO2(FITC)/PEI-FA can be exploited as a novel, non-viral, and concurrent targeting delivery system for targeted gene therapy as well as for MR imaging in cancer diagnosis.

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Field dependent magnetization at 25°C for Fe3O4@SiO2(FITC), Fe3O4@SiO2(FITC)/PEI-FA, and Fe3O4@SiO2(FITC)/PEI-FA/Notch-1 shRNA nanoparticles.
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f5: Field dependent magnetization at 25°C for Fe3O4@SiO2(FITC), Fe3O4@SiO2(FITC)/PEI-FA, and Fe3O4@SiO2(FITC)/PEI-FA/Notch-1 shRNA nanoparticles.

Mentions: FITC-APS conjugates were prepared in advance via an addition reaction between the isothiocyanate group of FITC dye and the primary amine group of APS. This synthetic process enables the co-encapsulation of Fe3O4 nanoparticles and a large number of FITC dye molecules to stay together within the silica shell42829. Using fluorescence microscopy, we confirmed that both Fe3O4@SiO2(FITC) and Fe3O4@SiO2(FITC)/PEI-FA nanoparticles were well-dispersed, and distinct fluorescence existed for the two nanoparticles (Figure 3). It indicated that PEI-FA modification had no obvious influence on fluorescence intensity (Figure 3B). The 1H NMR analysis was used to determine the composition of the resulting polymers. In the 1H-NMR spectrum of PEI-FA in D2O as shown in Figure 4A, the signals at δ 2.0–3.0 ppm were attributed to the methylene group in PEI, and the signals at δ 6.5–9.0 related to the H of benzene ring in FA3031. The results indicate that about one FA molecular chain was conjugated to one PEI molecular chain. To verify whether the PEI-FA co-polymer were coated on the surface of Fe3O4@SiO2(FITC) nanoparticles, fluorescence emission spectra were used and the results showed an emission peak at the wavelength of approximately 440 nm for the Fe3O4@SiO2(FITC)/PEI-FA nanoparticle suspension (Figure 4B), which is the characteristic fluorescence emission peak of FA41031, suggesting that PEI-FA molecules were successfully coated onto the surface of Fe3O4@SiO2(FITC) nanoparticles. The zeta potential analysis further showed a positive surface charge of +17.5 ± 1.3 mV (Figure 4C) for the Fe3O4@SiO2(FITC)/PEI-FA nanoparticle, suggesting that the Fe3O4@SiO2(FITC)/PEI-FA nanoparticle could bind to the negatively charged Notch-1 shRNA through electrostatic interaction. For assessment of the magnetic properties and sensitivity of the formulated nanoparticles, magnetic hysteresis loops were recorded using a magnetometer, and the three types of nanoparticles showed superparamagnetic behavior without magnetic hysteresis (Figure 5) at room temperature (about 300 K). They were found to have no coercive fields, thereby confirming their superparamagnetic nature. There was also no significant difference in their magnetic hysteresis loops and magnetization saturation values even when the Fe3O4@SiO2(FITC)/PEI-FA nanoparticles were modified with the Notch-1 shRNA.


Multifunctional core/shell nanoparticles cross-linked polyetherimide-folic acid as efficient Notch-1 siRNA carrier for targeted killing of breast cancer.

Yang H, Li Y, Li T, Xu M, Chen Y, Wu C, Dang X, Liu Y - Sci Rep (2014)

Field dependent magnetization at 25°C for Fe3O4@SiO2(FITC), Fe3O4@SiO2(FITC)/PEI-FA, and Fe3O4@SiO2(FITC)/PEI-FA/Notch-1 shRNA nanoparticles.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4233336&req=5

f5: Field dependent magnetization at 25°C for Fe3O4@SiO2(FITC), Fe3O4@SiO2(FITC)/PEI-FA, and Fe3O4@SiO2(FITC)/PEI-FA/Notch-1 shRNA nanoparticles.
Mentions: FITC-APS conjugates were prepared in advance via an addition reaction between the isothiocyanate group of FITC dye and the primary amine group of APS. This synthetic process enables the co-encapsulation of Fe3O4 nanoparticles and a large number of FITC dye molecules to stay together within the silica shell42829. Using fluorescence microscopy, we confirmed that both Fe3O4@SiO2(FITC) and Fe3O4@SiO2(FITC)/PEI-FA nanoparticles were well-dispersed, and distinct fluorescence existed for the two nanoparticles (Figure 3). It indicated that PEI-FA modification had no obvious influence on fluorescence intensity (Figure 3B). The 1H NMR analysis was used to determine the composition of the resulting polymers. In the 1H-NMR spectrum of PEI-FA in D2O as shown in Figure 4A, the signals at δ 2.0–3.0 ppm were attributed to the methylene group in PEI, and the signals at δ 6.5–9.0 related to the H of benzene ring in FA3031. The results indicate that about one FA molecular chain was conjugated to one PEI molecular chain. To verify whether the PEI-FA co-polymer were coated on the surface of Fe3O4@SiO2(FITC) nanoparticles, fluorescence emission spectra were used and the results showed an emission peak at the wavelength of approximately 440 nm for the Fe3O4@SiO2(FITC)/PEI-FA nanoparticle suspension (Figure 4B), which is the characteristic fluorescence emission peak of FA41031, suggesting that PEI-FA molecules were successfully coated onto the surface of Fe3O4@SiO2(FITC) nanoparticles. The zeta potential analysis further showed a positive surface charge of +17.5 ± 1.3 mV (Figure 4C) for the Fe3O4@SiO2(FITC)/PEI-FA nanoparticle, suggesting that the Fe3O4@SiO2(FITC)/PEI-FA nanoparticle could bind to the negatively charged Notch-1 shRNA through electrostatic interaction. For assessment of the magnetic properties and sensitivity of the formulated nanoparticles, magnetic hysteresis loops were recorded using a magnetometer, and the three types of nanoparticles showed superparamagnetic behavior without magnetic hysteresis (Figure 5) at room temperature (about 300 K). They were found to have no coercive fields, thereby confirming their superparamagnetic nature. There was also no significant difference in their magnetic hysteresis loops and magnetization saturation values even when the Fe3O4@SiO2(FITC)/PEI-FA nanoparticles were modified with the Notch-1 shRNA.

Bottom Line: Our results showed that Fe3O4@SiO2(FITC)/PEI-FA/Notch-1 shRNA nanoparticles are 64 nm in diameter with well dispersed and superparamagnetic.Magnetic resonance (MR) imaging and fluorescence microscopy showed significant preferential uptake of Fe3O4@SiO2(FITC)/PEI-FA/Notch-1 shRNA nanocomplex by MDA-MB-231 cells.Transfected MDA-MB-231 cells exhibited significantly decreased expression of Notch-1, inhibited cell proliferation, and increased cell apoptosis, leading to the killing of MDA-MB-231 cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Biophysics, School of Life Science and Technology, University of Electronic Science and Technology of China, Chengdu 610054, Sichuan, P.R. China.

ABSTRACT
In gene therapy, how genetic therapeutics can be efficiently and safely delivered into target tissues/cells remains a major obstacle to overcome. To address this issue, nanoparticles consisting of non-covalently coupled polyethyleneimine (PEI) and folic acid (FA) to the magnetic and fluorescent core/shell of Fe3O4@SiO2(FITC) was tested for their ability to deliver Notch-1 shRNA. Our results showed that Fe3O4@SiO2(FITC)/PEI-FA/Notch-1 shRNA nanoparticles are 64 nm in diameter with well dispersed and superparamagnetic. These nanoparticles with on significant cytotoxicity are capable of delivering Notch-1 shRNA into human breast cancer MDA-MB-231 cells with high efficiency while effectively protected shRNA from degradation by exogenous DNaseI and nucleases. Magnetic resonance (MR) imaging and fluorescence microscopy showed significant preferential uptake of Fe3O4@SiO2(FITC)/PEI-FA/Notch-1 shRNA nanocomplex by MDA-MB-231 cells. Transfected MDA-MB-231 cells exhibited significantly decreased expression of Notch-1, inhibited cell proliferation, and increased cell apoptosis, leading to the killing of MDA-MB-231 cells. In light of the magnetic targeting capabilities of Fe3O4@SiO2(FITC)/PEI-FA, our results show that by complexing with a second molecular targeting therapeutic, such as Notch-1 shRNA in this report, Fe3O4@SiO2(FITC)/PEI-FA can be exploited as a novel, non-viral, and concurrent targeting delivery system for targeted gene therapy as well as for MR imaging in cancer diagnosis.

Show MeSH
Related in: MedlinePlus