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Delivery of DNAzyme targeting aurora kinase A to inhibit the proliferation and migration of human prostate cancer.

Xing Z, Gao S, Duan Y, Han H, Li L, Yang Y, Li Q - Int J Nanomedicine (2015)

Bottom Line: Through flow cytometric analysis, an early apoptotic ratio of 25.93% and G2 phase of 22.58% has been detected after N-Ac-L-Leu-PEI-mediated DNAzyme transfection.Finally, wound healing and Transwell migration assay showed that DNAzyme transfection could efficiently inhibit the cell migration.These results demonstrated that N-Ac-L-Leu-PEI could successfully mediate the DNAzyme delivery and downregulate the expression level of aurora kinase A, triggering a significant inhibitory effect of excessive proliferation and migration of tumor cells.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory for Molecular Enzymology and Engineering of Ministry of Education, School of Life Sciences, Jilin University, Changchun, People's Republic of China.

ABSTRACT
Herein, a polyethylenimine derivative N-acetyl-L-leucine-polyethylenimine (N-Ac-L-Leu-PEI) was employed as a carrier to achieve the delivery of DNAzyme targeting aurora kinase A using PC-3 cell as a model. Flow cytometry and confocal laser scanning microscopy demonstrated that the derivative could realize the cellular uptake of nanoparticles in an energy-dependent and clathrin-mediated pathway and obtain a high DNAzyme concentration in the cytoplasm through further endosomal escape. After DNAzyme transfection, expression level of aurora kinase A would be downregulated at the protein level. Meanwhile, the inhibition of cell proliferation was observed through 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and cell colony formation assay, attributing to the activation of apoptosis and cell cycle arrest. Through flow cytometric analysis, an early apoptotic ratio of 25.93% and G2 phase of 22.58% has been detected after N-Ac-L-Leu-PEI-mediated DNAzyme transfection. Finally, wound healing and Transwell migration assay showed that DNAzyme transfection could efficiently inhibit the cell migration. These results demonstrated that N-Ac-L-Leu-PEI could successfully mediate the DNAzyme delivery and downregulate the expression level of aurora kinase A, triggering a significant inhibitory effect of excessive proliferation and migration of tumor cells.

No MeSH data available.


Related in: MedlinePlus

Gel retardation assay for the nanocomplexes of N-Ac-l-Leu-PEI with DNAzyme at different mass ratios.Abbreviation:N-Ac-l-Leu-PEI, N-acetyl-l-leucine-polyethylenimine.
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f1-ijn-10-5715: Gel retardation assay for the nanocomplexes of N-Ac-l-Leu-PEI with DNAzyme at different mass ratios.Abbreviation:N-Ac-l-Leu-PEI, N-acetyl-l-leucine-polyethylenimine.

Mentions: The derivative N-Ac-l-Leu-PEI was synthesized by 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide/N-hydroxysuccinimide-mediated couple reaction between PEI25K and hydrophobic N-Ac-l-Leu according to the previous research.31 The particle size and zeta potential of nanocomplex formed by N-Ac-l-Leu-PEI with DNAzyme at different mass ratios were determined (Table 1). The nanoparticles could be formed with a size range of 99–190 nm, and the particle size exhibited a decreasing tendency with an increasing ratio of polymer, indicating that DNAzyme was gradually packaged and condensed. Meanwhile, the nanoparticles formed by N-Ac-l-Leu-PEI and DNAzyme showed a positively charged state and zeta potential values increased with the increasing ratio of polymer, ranging from +15 mV to +23 mV. Thus, the particle size and zeta potential made these nanoparticles suitable for efficient endocytosis and further transfection. The binding affinity of N-Ac-l-Leu-PEI with DNAzyme was examined by agarose gel retardation assay (Figure 1). It was obviously observed that complete DNAzyme retardation could be achieved at a critical mass ratio of 2.0, indicating the formation of stable N-Ac-l-Leu-PEI/Dz nanopolyplex. Compared with plasmid DNA (critical mass ratio of 0.6–0.8),31N-Ac-l-Leu-PEI exhibited a relatively weaker binding ability for single-stranded oligoDNA segment.


Delivery of DNAzyme targeting aurora kinase A to inhibit the proliferation and migration of human prostate cancer.

Xing Z, Gao S, Duan Y, Han H, Li L, Yang Y, Li Q - Int J Nanomedicine (2015)

Gel retardation assay for the nanocomplexes of N-Ac-l-Leu-PEI with DNAzyme at different mass ratios.Abbreviation:N-Ac-l-Leu-PEI, N-acetyl-l-leucine-polyethylenimine.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4583550&req=5

f1-ijn-10-5715: Gel retardation assay for the nanocomplexes of N-Ac-l-Leu-PEI with DNAzyme at different mass ratios.Abbreviation:N-Ac-l-Leu-PEI, N-acetyl-l-leucine-polyethylenimine.
Mentions: The derivative N-Ac-l-Leu-PEI was synthesized by 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide/N-hydroxysuccinimide-mediated couple reaction between PEI25K and hydrophobic N-Ac-l-Leu according to the previous research.31 The particle size and zeta potential of nanocomplex formed by N-Ac-l-Leu-PEI with DNAzyme at different mass ratios were determined (Table 1). The nanoparticles could be formed with a size range of 99–190 nm, and the particle size exhibited a decreasing tendency with an increasing ratio of polymer, indicating that DNAzyme was gradually packaged and condensed. Meanwhile, the nanoparticles formed by N-Ac-l-Leu-PEI and DNAzyme showed a positively charged state and zeta potential values increased with the increasing ratio of polymer, ranging from +15 mV to +23 mV. Thus, the particle size and zeta potential made these nanoparticles suitable for efficient endocytosis and further transfection. The binding affinity of N-Ac-l-Leu-PEI with DNAzyme was examined by agarose gel retardation assay (Figure 1). It was obviously observed that complete DNAzyme retardation could be achieved at a critical mass ratio of 2.0, indicating the formation of stable N-Ac-l-Leu-PEI/Dz nanopolyplex. Compared with plasmid DNA (critical mass ratio of 0.6–0.8),31N-Ac-l-Leu-PEI exhibited a relatively weaker binding ability for single-stranded oligoDNA segment.

Bottom Line: Through flow cytometric analysis, an early apoptotic ratio of 25.93% and G2 phase of 22.58% has been detected after N-Ac-L-Leu-PEI-mediated DNAzyme transfection.Finally, wound healing and Transwell migration assay showed that DNAzyme transfection could efficiently inhibit the cell migration.These results demonstrated that N-Ac-L-Leu-PEI could successfully mediate the DNAzyme delivery and downregulate the expression level of aurora kinase A, triggering a significant inhibitory effect of excessive proliferation and migration of tumor cells.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory for Molecular Enzymology and Engineering of Ministry of Education, School of Life Sciences, Jilin University, Changchun, People's Republic of China.

ABSTRACT
Herein, a polyethylenimine derivative N-acetyl-L-leucine-polyethylenimine (N-Ac-L-Leu-PEI) was employed as a carrier to achieve the delivery of DNAzyme targeting aurora kinase A using PC-3 cell as a model. Flow cytometry and confocal laser scanning microscopy demonstrated that the derivative could realize the cellular uptake of nanoparticles in an energy-dependent and clathrin-mediated pathway and obtain a high DNAzyme concentration in the cytoplasm through further endosomal escape. After DNAzyme transfection, expression level of aurora kinase A would be downregulated at the protein level. Meanwhile, the inhibition of cell proliferation was observed through 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and cell colony formation assay, attributing to the activation of apoptosis and cell cycle arrest. Through flow cytometric analysis, an early apoptotic ratio of 25.93% and G2 phase of 22.58% has been detected after N-Ac-L-Leu-PEI-mediated DNAzyme transfection. Finally, wound healing and Transwell migration assay showed that DNAzyme transfection could efficiently inhibit the cell migration. These results demonstrated that N-Ac-L-Leu-PEI could successfully mediate the DNAzyme delivery and downregulate the expression level of aurora kinase A, triggering a significant inhibitory effect of excessive proliferation and migration of tumor cells.

No MeSH data available.


Related in: MedlinePlus