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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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(A) Cellular internalization of nanoparticles into human breast cancer MDA-MB-231 cells was observed by an inverted microscope after 6 h incubation with Fe3O4@SiO2(FITC) (NPs), Fe3O4@SiO2(FITC)/PEI (NPs/PEI), Fe3O4@SiO2(FITC)/PEI-FA (NPs/PEI-FA), and FA pretreated MDA-MB-231 cells before Fe3O4@SiO2(FITC)/PEI-FA addition (FA + NPs/PEI-FA). (B) Quantitative cellular uptake of the nanoparticles into MDA-MB-231 cells through measuring mean fluorescence intensity of FITC fluorescence in the cells, which was normalized to the control (NPs) (n = 5, mean ± SD). *p < 0.05 compared with NPs, **p < 0.01 compared with NPs, #p < 0.05 compared with NPs/PEI-FA.
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f9: (A) Cellular internalization of nanoparticles into human breast cancer MDA-MB-231 cells was observed by an inverted microscope after 6 h incubation with Fe3O4@SiO2(FITC) (NPs), Fe3O4@SiO2(FITC)/PEI (NPs/PEI), Fe3O4@SiO2(FITC)/PEI-FA (NPs/PEI-FA), and FA pretreated MDA-MB-231 cells before Fe3O4@SiO2(FITC)/PEI-FA addition (FA + NPs/PEI-FA). (B) Quantitative cellular uptake of the nanoparticles into MDA-MB-231 cells through measuring mean fluorescence intensity of FITC fluorescence in the cells, which was normalized to the control (NPs) (n = 5, mean ± SD). *p < 0.05 compared with NPs, **p < 0.01 compared with NPs, #p < 0.05 compared with NPs/PEI-FA.

Mentions: In vitro cellular uptake experiments were performed using human breast carcinoma MDA-MB-231 cells, known to significantly overexpress the folate receptor152526. The uptake of the nanoparticle conjugates was evaluated after 6-h cell culture, and is shown in Figure 9. In contrast to Fe3O4@SiO2(FITC) or Fe3O4@SiO2(FITC)/PEI, the uptake of FA-conjugated Fe3O4@SiO2(FITC)/PEI-FA by MDA-MB-231 cells was ~3-fold higher than those cultured with Fe3O4@SiO2(FITC). PEI, a cationic polymer, could also facilitate nanoparticle uptake in some extent. While the increase in intracellular Fe3O4@SiO2(FITC)/PEI nanoparticles was somewhat visible, however, the fluorescent intensity was considerably less than that in FA-conjugated Fe3O4@SiO2(FITC)/PEI-FA nanoparticles. Furthermore, high uptake was also observed in Fe3O4@SiO2(FITC)/PEI-FA/Notch-1 shRNA nanocomplex (Figure 10), which demonstrated that absorption of Notch-1 shRNA to the nanoparticles did not affect uptake. To ascertain that the nanoparticles were indeed internalized rather than being bound to cell membrane, MDA-MB-231 cells incubated with various nanoparticles were examined by confocal fluorescence microscopy. As shown in Figure 10, the nanoparticles mainly localized to the cytoplasm after 6 h culture. These results agreed well with that of inverted fluorescence microscopy, and also confirmed that the uptake of FA-conjugated Fe3O4@SiO2(FITC)/PEI-FA nanoparticles was more favorable than other non-FA-conjugated nanoparticles.


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)

(A) Cellular internalization of nanoparticles into human breast cancer MDA-MB-231 cells was observed by an inverted microscope after 6 h incubation with Fe3O4@SiO2(FITC) (NPs), Fe3O4@SiO2(FITC)/PEI (NPs/PEI), Fe3O4@SiO2(FITC)/PEI-FA (NPs/PEI-FA), and FA pretreated MDA-MB-231 cells before Fe3O4@SiO2(FITC)/PEI-FA addition (FA + NPs/PEI-FA). (B) Quantitative cellular uptake of the nanoparticles into MDA-MB-231 cells through measuring mean fluorescence intensity of FITC fluorescence in the cells, which was normalized to the control (NPs) (n = 5, mean ± SD). *p < 0.05 compared with NPs, **p < 0.01 compared with NPs, #p < 0.05 compared with NPs/PEI-FA.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f9: (A) Cellular internalization of nanoparticles into human breast cancer MDA-MB-231 cells was observed by an inverted microscope after 6 h incubation with Fe3O4@SiO2(FITC) (NPs), Fe3O4@SiO2(FITC)/PEI (NPs/PEI), Fe3O4@SiO2(FITC)/PEI-FA (NPs/PEI-FA), and FA pretreated MDA-MB-231 cells before Fe3O4@SiO2(FITC)/PEI-FA addition (FA + NPs/PEI-FA). (B) Quantitative cellular uptake of the nanoparticles into MDA-MB-231 cells through measuring mean fluorescence intensity of FITC fluorescence in the cells, which was normalized to the control (NPs) (n = 5, mean ± SD). *p < 0.05 compared with NPs, **p < 0.01 compared with NPs, #p < 0.05 compared with NPs/PEI-FA.
Mentions: In vitro cellular uptake experiments were performed using human breast carcinoma MDA-MB-231 cells, known to significantly overexpress the folate receptor152526. The uptake of the nanoparticle conjugates was evaluated after 6-h cell culture, and is shown in Figure 9. In contrast to Fe3O4@SiO2(FITC) or Fe3O4@SiO2(FITC)/PEI, the uptake of FA-conjugated Fe3O4@SiO2(FITC)/PEI-FA by MDA-MB-231 cells was ~3-fold higher than those cultured with Fe3O4@SiO2(FITC). PEI, a cationic polymer, could also facilitate nanoparticle uptake in some extent. While the increase in intracellular Fe3O4@SiO2(FITC)/PEI nanoparticles was somewhat visible, however, the fluorescent intensity was considerably less than that in FA-conjugated Fe3O4@SiO2(FITC)/PEI-FA nanoparticles. Furthermore, high uptake was also observed in Fe3O4@SiO2(FITC)/PEI-FA/Notch-1 shRNA nanocomplex (Figure 10), which demonstrated that absorption of Notch-1 shRNA to the nanoparticles did not affect uptake. To ascertain that the nanoparticles were indeed internalized rather than being bound to cell membrane, MDA-MB-231 cells incubated with various nanoparticles were examined by confocal fluorescence microscopy. As shown in Figure 10, the nanoparticles mainly localized to the cytoplasm after 6 h culture. These results agreed well with that of inverted fluorescence microscopy, and also confirmed that the uptake of FA-conjugated Fe3O4@SiO2(FITC)/PEI-FA nanoparticles was more favorable than other non-FA-conjugated nanoparticles.

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