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PLGA-based gene delivering nanoparticle enhance suppression effect of miRNA in HePG2 cells.

Liang GF, Zhu YL, Sun B, Hu FH, Tian T, Li SC, Xiao ZD - Nanoscale Res Lett (2011)

Bottom Line: The N/P ratio (ratio of the polymer nitrogen to the DNA phosphate) 6 of the PLGA/PEI/DNA nanocomplex displays the best property among various N/P proportions, yielding similar transfection efficiency when compared to Lipofectamine/DNA lipoplexes.Moreover, nanocomplex shows better serum compatibility than commercial liposome.Therefore, these results demonstrate that PLGA/PEI nanoparticles are promising non-viral vectors for gene delivery.

View Article: PubMed Central - HTML - PubMed

Affiliation: State Key Laboratory of Bioelectronics, School of Biological Science and Medical Engineering, Southeast University, Nanjing, 210096, China. zdxiao@seu.edu.

ABSTRACT
The biggest challenge in the field of gene therapy is how to effectively deliver target genes to special cells. This study aimed to develop a new type of poly(D,L-lactide-co-glycolide) (PLGA)-based nanoparticles for gene delivery, which are capable of overcoming the disadvantages of polyethylenimine (PEI)- or cationic liposome-based gene carrier, such as the cytotoxicity induced by excess positive charge, as well as the aggregation on the cell surface. The PLGA-based nanoparticles presented in this study were synthesized by emulsion evaporation method and characterized by transmission electron microscopy, dynamic light scattering, and energy dispersive spectroscopy. The size of PLGA/PEI nanoparticles in phosphate-buffered saline (PBS) was about 60 nm at the optimal charge ratio. Without observable aggregation, the nanoparticles showed a better monodispersity. The PLGA-based nanoparticles were used as vector carrier for miRNA transfection in HepG2 cells. It exhibited a higher transfection efficiency and lower cytotoxicity in HepG2 cells compared to the PEI/DNA complex. The N/P ratio (ratio of the polymer nitrogen to the DNA phosphate) 6 of the PLGA/PEI/DNA nanocomplex displays the best property among various N/P proportions, yielding similar transfection efficiency when compared to Lipofectamine/DNA lipoplexes. Moreover, nanocomplex shows better serum compatibility than commercial liposome. PLGA nanocomplexes obviously accumulate in tumor cells after transfection, which indicate that the complexes contribute to cellular uptake of pDNA and pronouncedly enhance the treatment effect of miR-26a by inducing cell cycle arrest. Therefore, these results demonstrate that PLGA/PEI nanoparticles are promising non-viral vectors for gene delivery.

No MeSH data available.


Related in: MedlinePlus

Relative expression change of miR-26a level. (1) Untransfected group; (2) naked DNA group; (3) PLGA/PEI transfected group. P < 0.05 compared with groups naked DNA and PLGA/PEI nanoparticles.
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Figure 5: Relative expression change of miR-26a level. (1) Untransfected group; (2) naked DNA group; (3) PLGA/PEI transfected group. P < 0.05 compared with groups naked DNA and PLGA/PEI nanoparticles.

Mentions: To examine the biological activities of miR-26a delivered by nanoparticles/pDNA in HepG2 cells, the expression levels of miR-26a were detected by qRT-PCR in transfected HepG2 cells. Forty-eight hours after transfection, HepG2 cells were harvested and total RNA was extracted for monitoring miRNA expression. Real-time quantitative RT-PCR results show that the miR-26a level in the transfected cells is increased by 7.73-folds compared with untransfected cells (p < 0.05) (Figure 5). In contrast, the expression level of miR-26a shows no obvious change in cells transfected with negative control (miR-NC) and naked pDNA (p > 0.05).


PLGA-based gene delivering nanoparticle enhance suppression effect of miRNA in HePG2 cells.

Liang GF, Zhu YL, Sun B, Hu FH, Tian T, Li SC, Xiao ZD - Nanoscale Res Lett (2011)

Relative expression change of miR-26a level. (1) Untransfected group; (2) naked DNA group; (3) PLGA/PEI transfected group. P < 0.05 compared with groups naked DNA and PLGA/PEI nanoparticles.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3211866&req=5

Figure 5: Relative expression change of miR-26a level. (1) Untransfected group; (2) naked DNA group; (3) PLGA/PEI transfected group. P < 0.05 compared with groups naked DNA and PLGA/PEI nanoparticles.
Mentions: To examine the biological activities of miR-26a delivered by nanoparticles/pDNA in HepG2 cells, the expression levels of miR-26a were detected by qRT-PCR in transfected HepG2 cells. Forty-eight hours after transfection, HepG2 cells were harvested and total RNA was extracted for monitoring miRNA expression. Real-time quantitative RT-PCR results show that the miR-26a level in the transfected cells is increased by 7.73-folds compared with untransfected cells (p < 0.05) (Figure 5). In contrast, the expression level of miR-26a shows no obvious change in cells transfected with negative control (miR-NC) and naked pDNA (p > 0.05).

Bottom Line: The N/P ratio (ratio of the polymer nitrogen to the DNA phosphate) 6 of the PLGA/PEI/DNA nanocomplex displays the best property among various N/P proportions, yielding similar transfection efficiency when compared to Lipofectamine/DNA lipoplexes.Moreover, nanocomplex shows better serum compatibility than commercial liposome.Therefore, these results demonstrate that PLGA/PEI nanoparticles are promising non-viral vectors for gene delivery.

View Article: PubMed Central - HTML - PubMed

Affiliation: State Key Laboratory of Bioelectronics, School of Biological Science and Medical Engineering, Southeast University, Nanjing, 210096, China. zdxiao@seu.edu.

ABSTRACT
The biggest challenge in the field of gene therapy is how to effectively deliver target genes to special cells. This study aimed to develop a new type of poly(D,L-lactide-co-glycolide) (PLGA)-based nanoparticles for gene delivery, which are capable of overcoming the disadvantages of polyethylenimine (PEI)- or cationic liposome-based gene carrier, such as the cytotoxicity induced by excess positive charge, as well as the aggregation on the cell surface. The PLGA-based nanoparticles presented in this study were synthesized by emulsion evaporation method and characterized by transmission electron microscopy, dynamic light scattering, and energy dispersive spectroscopy. The size of PLGA/PEI nanoparticles in phosphate-buffered saline (PBS) was about 60 nm at the optimal charge ratio. Without observable aggregation, the nanoparticles showed a better monodispersity. The PLGA-based nanoparticles were used as vector carrier for miRNA transfection in HepG2 cells. It exhibited a higher transfection efficiency and lower cytotoxicity in HepG2 cells compared to the PEI/DNA complex. The N/P ratio (ratio of the polymer nitrogen to the DNA phosphate) 6 of the PLGA/PEI/DNA nanocomplex displays the best property among various N/P proportions, yielding similar transfection efficiency when compared to Lipofectamine/DNA lipoplexes. Moreover, nanocomplex shows better serum compatibility than commercial liposome. PLGA nanocomplexes obviously accumulate in tumor cells after transfection, which indicate that the complexes contribute to cellular uptake of pDNA and pronouncedly enhance the treatment effect of miR-26a by inducing cell cycle arrest. Therefore, these results demonstrate that PLGA/PEI nanoparticles are promising non-viral vectors for gene delivery.

No MeSH data available.


Related in: MedlinePlus