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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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Tumor volume of RCC-bearing nude mice treated with the FA-PEAs:VHL polyplexes. The tumor size in each group was the mean value of six nude mice. A value of p less than 0.05 was accepted to be significant. *, p < 0.05. All the data were presented as mean ± SEM (standard error of mean), and the statistics were performed with Student's t test.
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Figure 4: Tumor volume of RCC-bearing nude mice treated with the FA-PEAs:VHL polyplexes. The tumor size in each group was the mean value of six nude mice. A value of p less than 0.05 was accepted to be significant. *, p < 0.05. All the data were presented as mean ± SEM (standard error of mean), and the statistics were performed with Student's t test.

Mentions: Based on the transfection effects of three modified PEI in vitro, we further tested whether the PEI-derived nanoparticles could efficiently transfer target gene into tumor tissues in vivo. The RCC tumor xenograft was established by inoculation with OS-RC-2 cells in nude mice to verify VHL gene therapy effects. Compared with the control group, the FA-PEAs:pVHL group obviously exhibited anti-tumor activity in the tumor-bearing nude mice from 21 to 31 days after implantation with OS-RC-2 cells. On day 31 post-implantation, the mean tumor volume was 678.70 ± 121.73 mm3 in FA-PEAs:pVHL-treated mice, whereas tumor had 935.23 ± 112.14, 950.40 ± 41.98 mm3 in FA-PEAs:pVITRO2-treated and PBS group respectively (Figure 4, p < 0.05), which indicated VHL gene therapy achieved about 30% inhibiting rate of tumor growth. Whereas there was no statistical difference between the PBS group and the FA-PEAs:pVITRO2-treated group in tumor size. These data indicated that the FA-PEAs:pVHL therapy can inhibit tumor growth in vivo.


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)

Tumor volume of RCC-bearing nude mice treated with the FA-PEAs:VHL polyplexes. The tumor size in each group was the mean value of six nude mice. A value of p less than 0.05 was accepted to be significant. *, p < 0.05. All the data were presented as mean ± SEM (standard error of mean), and the statistics were performed with Student's t test.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 4: Tumor volume of RCC-bearing nude mice treated with the FA-PEAs:VHL polyplexes. The tumor size in each group was the mean value of six nude mice. A value of p less than 0.05 was accepted to be significant. *, p < 0.05. All the data were presented as mean ± SEM (standard error of mean), and the statistics were performed with Student's t test.
Mentions: Based on the transfection effects of three modified PEI in vitro, we further tested whether the PEI-derived nanoparticles could efficiently transfer target gene into tumor tissues in vivo. The RCC tumor xenograft was established by inoculation with OS-RC-2 cells in nude mice to verify VHL gene therapy effects. Compared with the control group, the FA-PEAs:pVHL group obviously exhibited anti-tumor activity in the tumor-bearing nude mice from 21 to 31 days after implantation with OS-RC-2 cells. On day 31 post-implantation, the mean tumor volume was 678.70 ± 121.73 mm3 in FA-PEAs:pVHL-treated mice, whereas tumor had 935.23 ± 112.14, 950.40 ± 41.98 mm3 in FA-PEAs:pVITRO2-treated and PBS group respectively (Figure 4, p < 0.05), which indicated VHL gene therapy achieved about 30% inhibiting rate of tumor growth. Whereas there was no statistical difference between the PBS group and the FA-PEAs:pVITRO2-treated group in tumor size. These data indicated that the FA-PEAs:pVHL therapy can inhibit tumor growth in vivo.

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