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Comparisons of three polyethyleneimine-derived nanoparticles as a gene therapy delivery system for renal cell carcinoma.

Xu Z, Shen G, Xia X, Zhao X, Zhang P, Wu H, Guo Q, Qian Z, Wei Y, Liang S - J Transl Med (2011)

Bottom Line: Each of three modified PEI-derived biomaterials had an increased transfection efficiency and a lower cytotoxicity compared with its precursor PEI with 25-kD or 2-kD molecule weight in vitro.And the mean tumor volume was obviously decreased 30% by using FA-PEAs to transfer VHL plasmids to treat mice RCC models.The VHL gene expression was greatly improved in the VHL-treated group.

View Article: PubMed Central - HTML - PubMed

Affiliation: State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, West China Medical School, Sichuan University, Chengdu, P. R. China

ABSTRACT

Background: Polyethyleneimine (PEI), which can interact with negatively charged DNA through electrostatic interaction to form nanocomplexes, has been widely attempted to use as a gene delivery system. However, PEI has some defects that are not fit for keeping on gene expression. Therefore, some modifications against PEI properties have been done to improve their application value in gene delivery. In this study, three modified PEI derivatives, including poly(ε-caprolactone)-pluronic-poly(ε-caprolactone) grafted PEI (PCFC-g-PEI), folic acid-PCFC-isophorone diidocyanate-PEI (FA-PEAs) and heparin-PEI (HPEI), were evaluated in terms of their cytotoxicity and transfection efficiency in vitro and in vivo in order to ascertain their potential application in gene therapy.

Methods: MTT assay and a marker GFP gene, encoding green fluorescent protein, were used to evaluate cell toxicity and transfection activity of the three modified PEI in vitro. Renal cell carcinoma (RCC) models were established in BALB/c nude mice inoculated with OS-RC-2 cells to detect the gene therapy effects using the three PEI-derived nanoparticles as gene delivery vehicles. The expression status of a target gene Von Hippel-Lindau (VHL) in treated tumor tissues was analyzed by semiquantitative RT-PCR and immunohistochemistry.

Results: Each of three modified PEI-derived biomaterials had an increased transfection efficiency and a lower cytotoxicity compared with its precursor PEI with 25-kD or 2-kD molecule weight in vitro. And the mean tumor volume was obviously decreased 30% by using FA-PEAs to transfer VHL plasmids to treat mice RCC models. The VHL gene expression was greatly improved in the VHL-treated group. While there was no obvious tumor inhibition treated by PCFC-g-PEI:VHL and HPEI:VHL complexes.

Conclusions: The three modified PEI-derived biomaterials, including PCFC-g-PEI, FA-PEAs and HPEI, had an increased transfection efficiency in vitro and obviously lower toxicities compared with their precursor PEI molecules. The FA-PEAs probably provide a potential gene delivery system to treat RCC even other cancers in future.

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Transfection efficiency for GFP plasmid in OS-RC-2 cells mediated by the PEI and PEI-derived nanoparticles. The Figure 2A was shown the transfection effects of PCFC-g-PEI:GFP polyplexes at different weight ratios versus the control PEI (25 kD):GFP group. The transfection efficiency of FA-PEAs:GFP and HPEI:GFP complexes was respectively compared with the PEI (2-kD)/GFP (Figure 2B). The error bars represented the standard deviation of experimental repeats.
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Figure 3: Transfection efficiency for GFP plasmid in OS-RC-2 cells mediated by the PEI and PEI-derived nanoparticles. The Figure 2A was shown the transfection effects of PCFC-g-PEI:GFP polyplexes at different weight ratios versus the control PEI (25 kD):GFP group. The transfection efficiency of FA-PEAs:GFP and HPEI:GFP complexes was respectively compared with the PEI (2-kD)/GFP (Figure 2B). The error bars represented the standard deviation of experimental repeats.

Mentions: The GFP plasmid was used as a marker gene to detect cell transfection activity in vitro. The Figure 2 was the representative fluorescence profiling of positive GFP-expressing cells transfected with three PEI-derived nanoparticles. For PCFC-g-PEI, the average transfection efficiency was 12.1 ± 1.5%, 27.0 ± 2.1% and 7.9 ± 1.8% corresponding to 0.7:1, 1:1 and 1.3:1 ratio of PCFC-g-PEI:GFP complex (Figure 3A). While under same conditions, the transfection rate mediated with 25-kD PEI was 22.6 ± 1.7%, lower than 27.0 ± 2.1% which was transfected with PCFC-g-PEI:GFP at 1:1 weight ratio. Therefore, when the weight ratio of PCFC-g-PEI: GFP was 1, meanwhile, the concentration of PCFC-g-PEI was 2 μg/ml, the transfection rate of PCFC-g-PEI was highest and had no apparent toxicity on cells (<10 μg/ml). Thus, PCFC-g-PEI, with the 1:1 optimized weight ratio, had a higher transfection activity (Figure 2A-2B, Figure 3A) and a lower toxicity when compared to 25-kD PEI.


Comparisons of three polyethyleneimine-derived nanoparticles as a gene therapy delivery system for renal cell carcinoma.

Xu Z, Shen G, Xia X, Zhao X, Zhang P, Wu H, Guo Q, Qian Z, Wei Y, Liang S - J Transl Med (2011)

Transfection efficiency for GFP plasmid in OS-RC-2 cells mediated by the PEI and PEI-derived nanoparticles. The Figure 2A was shown the transfection effects of PCFC-g-PEI:GFP polyplexes at different weight ratios versus the control PEI (25 kD):GFP group. The transfection efficiency of FA-PEAs:GFP and HPEI:GFP complexes was respectively compared with the PEI (2-kD)/GFP (Figure 2B). The error bars represented the standard deviation of experimental repeats.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3108928&req=5

Figure 3: Transfection efficiency for GFP plasmid in OS-RC-2 cells mediated by the PEI and PEI-derived nanoparticles. The Figure 2A was shown the transfection effects of PCFC-g-PEI:GFP polyplexes at different weight ratios versus the control PEI (25 kD):GFP group. The transfection efficiency of FA-PEAs:GFP and HPEI:GFP complexes was respectively compared with the PEI (2-kD)/GFP (Figure 2B). The error bars represented the standard deviation of experimental repeats.
Mentions: The GFP plasmid was used as a marker gene to detect cell transfection activity in vitro. The Figure 2 was the representative fluorescence profiling of positive GFP-expressing cells transfected with three PEI-derived nanoparticles. For PCFC-g-PEI, the average transfection efficiency was 12.1 ± 1.5%, 27.0 ± 2.1% and 7.9 ± 1.8% corresponding to 0.7:1, 1:1 and 1.3:1 ratio of PCFC-g-PEI:GFP complex (Figure 3A). While under same conditions, the transfection rate mediated with 25-kD PEI was 22.6 ± 1.7%, lower than 27.0 ± 2.1% which was transfected with PCFC-g-PEI:GFP at 1:1 weight ratio. Therefore, when the weight ratio of PCFC-g-PEI: GFP was 1, meanwhile, the concentration of PCFC-g-PEI was 2 μg/ml, the transfection rate of PCFC-g-PEI was highest and had no apparent toxicity on cells (<10 μg/ml). Thus, PCFC-g-PEI, with the 1:1 optimized weight ratio, had a higher transfection activity (Figure 2A-2B, Figure 3A) and a lower toxicity when compared to 25-kD PEI.

Bottom Line: Each of three modified PEI-derived biomaterials had an increased transfection efficiency and a lower cytotoxicity compared with its precursor PEI with 25-kD or 2-kD molecule weight in vitro.And the mean tumor volume was obviously decreased 30% by using FA-PEAs to transfer VHL plasmids to treat mice RCC models.The VHL gene expression was greatly improved in the VHL-treated group.

View Article: PubMed Central - HTML - PubMed

Affiliation: State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, West China Medical School, Sichuan University, Chengdu, P. R. China

ABSTRACT

Background: Polyethyleneimine (PEI), which can interact with negatively charged DNA through electrostatic interaction to form nanocomplexes, has been widely attempted to use as a gene delivery system. However, PEI has some defects that are not fit for keeping on gene expression. Therefore, some modifications against PEI properties have been done to improve their application value in gene delivery. In this study, three modified PEI derivatives, including poly(ε-caprolactone)-pluronic-poly(ε-caprolactone) grafted PEI (PCFC-g-PEI), folic acid-PCFC-isophorone diidocyanate-PEI (FA-PEAs) and heparin-PEI (HPEI), were evaluated in terms of their cytotoxicity and transfection efficiency in vitro and in vivo in order to ascertain their potential application in gene therapy.

Methods: MTT assay and a marker GFP gene, encoding green fluorescent protein, were used to evaluate cell toxicity and transfection activity of the three modified PEI in vitro. Renal cell carcinoma (RCC) models were established in BALB/c nude mice inoculated with OS-RC-2 cells to detect the gene therapy effects using the three PEI-derived nanoparticles as gene delivery vehicles. The expression status of a target gene Von Hippel-Lindau (VHL) in treated tumor tissues was analyzed by semiquantitative RT-PCR and immunohistochemistry.

Results: Each of three modified PEI-derived biomaterials had an increased transfection efficiency and a lower cytotoxicity compared with its precursor PEI with 25-kD or 2-kD molecule weight in vitro. And the mean tumor volume was obviously decreased 30% by using FA-PEAs to transfer VHL plasmids to treat mice RCC models. The VHL gene expression was greatly improved in the VHL-treated group. While there was no obvious tumor inhibition treated by PCFC-g-PEI:VHL and HPEI:VHL complexes.

Conclusions: The three modified PEI-derived biomaterials, including PCFC-g-PEI, FA-PEAs and HPEI, had an increased transfection efficiency in vitro and obviously lower toxicities compared with their precursor PEI molecules. The FA-PEAs probably provide a potential gene delivery system to treat RCC even other cancers in future.

Show MeSH
Related in: MedlinePlus